Compositions and methods for inhibition of the jak pathway

ABSTRACT

The invention encompasses compounds having formula I and the compositions and methods using these compounds in the treatment of conditions in which modulation of the JAK pathway or inhibition of JAK kinases, particularly JAK3, are therapeutically useful.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of the earlier filing date of U.S.provisional application Ser. No. 61/140,604, filed Dec. 23, 2008.

I. INTRODUCTION

A. Field

The present invention relates to compounds, prodrugs, and methods ofusing these compounds and prodrugs thereof in the treatment ofconditions in which modulation of the JAK pathway or inhibition of JAKkinases, particularly JAK3, are therapeutically useful.

B. Background

Protein kinases constitute a large family of structurally relatedenzymes that are responsible for the control of a variety of signaltransduction processes within cells (see, e.g., Hardie and Hanks, TheProtein Kinase Facts Book, I and II, Academic Press, San Diego, Calif.,1995). Protein kinases are thought to have evolved from a commonancestral gene due to the conservation of their structure and catalyticfunction. Almost all kinases contain a similar 250-300 amino acidcatalytic domain. The kinases can be categorized into families by thesubstrates they phosphorylate (e.g., protein-tyrosine,protein-serine/threonine, lipids, etc.). Sequence motifs have beenidentified that generally correspond to each of these families (see,e.g., Hanks & Hunter, (1995), FASEB J. 9:576-596; Knighton et al.,(1991), Science 253:407-414; Hiles et al., (1992), Cell 70:419-429; Kunzet al., (1993), Cell 73:585-596; Garcia-Bustos et al., (1994), EMBO J.13:2352-2361).

JAK kinases (JAnus Kinases) are a family of cytoplasmic protein tyrosinekinases including JAK1, JAK2, JAK3 and TYK2. Each of the JAK kinases isselective for the receptors of certain cytokines, though multiple JAKkinases can be affected by particular cytokine or signaling pathways.Studies suggest that JAK3 associates with the common gamma (γc) chain ofthe various cytokine receptors. JAK3 in particular selectively binds toreceptors and is part of the cytokine signaling pathway for IL-2, IL-4,IL-7, IL-9, IL-15 and IL-21. JAK1 interacts with, among others, thereceptors for cytokines IL-2, IL-4, IL-7, IL-9 and IL-21, while JAK2interacts with, among others, the receptors for IL-9 and TNF-α. Upon thebinding of certain cytokines to their receptors (e.g., IL-2, IL-4, IL-7,IL-9, IL-15 and IL-21), receptor oligomerization occurs, resulting inthe cytoplasmic tails of associated JAK kinases being brought intoproximity and facilitating the trans-phosphorylation of tyrosineresidues on the JAK kinase. This trans-phosphorylation results in theactivation of the JAK kinase.

Phosphorylated JAK kinases bind various STAT (Signal Transducer andActivator of Transcription) proteins. STAT proteins, which are DNAbinding proteins activated by phosphorylation of tyrosine residues,function both as signaling molecules and transcription factors andultimately bind to specific DNA sequences present in the promoters ofcytokine-responsive genes (Leonard et al., (2000), J. Allergy Clin.Immunol. 105:877-888). JAK/STAT signaling has been implicated in themediation of many abnormal immune responses such as allergies, asthma,autoimmune diseases such as transplant (allograft) rejection, rheumatoidarthritis, amyotrophic lateral sclerosis and multiple sclerosis, as wellas in solid and hematologic malignancies such as leukemia and lymphomas.For a review of the pharmaceutical intervention of the JAK/STAT pathwaysee Frank, (1999), Mol. Med. 5:432:456 and Seidel et al., (2000),Oncogene 19:2645-2656.

JAK3 in particular has been implicated in a variety of biologicalprocesses. For example, the proliferation and survival of murine mastcells induced by IL-4 and IL-9 have been shown to be dependent on JAK3-and gamma chain-signaling (Suzuki et al., (2000), Blood 96:2172-2180).JAK3 also plays a crucial role in IgE receptor—mediated mast celldegranulation responses (Malaviya et al., (1999), Biochem. Biophys. Res.Commun. 257:807-813), and inhibition of JAK3 kinase has been shown toprevent type I hypersensitivity reactions, including anaphylaxis(Malaviya et al., (1999), J. Biol. Chem. 274:27028-27038). JAK3inhibition has also been shown to result in immune suppression forallograft rejection (Kirken, (2001), Transpl. Proc. 33:3268-3270). JAK3kinases have also been implicated in the mechanism involved in early andlate stages of rheumatoid arthritis (Muller-Ladner et al., (2000), J.Immunal. 164:3894-3901); familial amyotrophic lateral sclerosis (Trieuet al., (2000), Biochem Biophys. Res. Commun. 267:22-25); leukemia(Sudbeck et al., (1999), Clin. Cancer Res. 5:1569-1582); mycosisfungoides, a form of T-cell lymphoma (Nielsen et al., (1997), Prac.Natl. Acad. Sci. USA 94:6764-6769); and abnormal cell growth (Yu et al.,(1997), J. Immunol. 159:5206-5210; Catlett-Falcone et al., (1999),Immunity 10:105-115).

The JAK kinases, including JAK3, are abundantly expressed in primaryleukemic cells from children with acute lymphoblastic leukemia, the mostcommon form of childhood cancer, and studies have correlated STATactivation in certain cells with signals regulating apoptosis (Demoulinet al., (1996), Mol. Cell. Biol. 16:4710-6; Jurlander et al., (1997),Blood. 89:4146-52; Kaneko et al., (1997), Clin. Exp. Immun. 109:185-193;and Nakamura et al., (1996), J. Biol. Chem. 271:19483-8). They are alsoknown to be important to lymphocyte differentiation, function andsurvival. JAK-3 in particular plays an essential role in the function oflymphocytes, macrophages, and mast cells. Given the importance of thisJAK kinase, compounds which modulate the JAK pathway, including thoseselective for JAK3, can be useful for treating diseases or conditionswhere the function of lymphocytes, macrophages, or mast cells isinvolved (Kudlacz et al., (2004) Am. J. Transplant 4:51-57; Changelian(2003) Science 302:875-878). Conditions in which targeting of the JAKpathway or modulation of the JAK kinases, particularly JAK3, arecontemplated to be therapeutically useful include, leukemia, lymphoma,transplant rejection (e.g., pancreas islet transplant rejection, bonemarrow transplant applications (e.g., graft-versus-host disease),autoimmune diseases (e.g., diabetes), and inflammation (e.g., asthma,allergic reactions). Conditions which can benefit for inhibition of JAK3are discussed in greater detail below.

In view of the numerous conditions that are contemplated to benefit bytreatment involving modulation of the JAK pathway it is immediatelyapparent that new compounds that modulate JAK pathways and methods ofusing these compounds should provide substantial therapeutic benefits toa wide variety of patients. Provided herein are novel2,4-pyrimidinediamine compounds for use in the treatment of conditionsin which targeting of the JAK pathway or inhibition of JAK kinases,particularly JAK3, are therapeutically useful.

II. SUMMARY

The invention is directed to compounds, prodrugs, pharmaceuticalcompositions and methods of using them in the treatment of conditions inwhich modulation of the JAK pathway or inhibition of JAK kinases,particularly JAK3, will be therapeutically useful.

In one embodiment, this invention provides a compound of formula I:

wherein:

-   -   p is 0, 1, 2, or 3;    -   q is 0, 1, 2, or 3;    -   X is hydrogen, alkyl, substituted alkyl, hydroxy, alkoxy,        substituted alkoxy, amino, substituted amino, carboxyl, carboxyl        ester, —C(O)N(R⁴)R⁵, cyano, halo, nitro, alkenyl, substituted        alkenyl, alkynyl, substituted alkynyl, cycloalkynyl or        substituted cycloalkynyl;    -   Y is a straight or branched chain C₁₋₆ alkylene group,        cycloalkylene or substituted cycloalkylene;    -   R¹ is hydrogen, alkyl, substituted alkyl, cycloalkyl or        substituted cycloalkyl;    -   each R² independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, amino, substituted amino, aryl, substituted        aryl, aryloxy, substituted aryloxy, cyano, cycloalkyl,        substituted cycloalkyl, cycloalkoxy, substituted cycloalkoxy,        heteroaryl, substituted heteroaryl, heteroaryloxy, substituted        heteroaryloxy, heterocyclyl, substituted heterocyclyl,        heterocyclyloxy, substituted heterocyclyloxy, aminoacyl,        aminoacyloxy, carboxyl, carboxyl ester, carbonate ester,        —C(O)N(R⁴)R⁵, nitro or halo;    -   each R³ independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, amino, substituted amino, aryl, substituted        aryl, aryloxy, substituted aryloxy, cyano, cycloalkyl,        substituted cycloalkyl, cycloalkoxy, substituted cycloalkoxy,        heteroaryl, substituted heteroaryl, heteroaryloxy, substituted        heteroaryloxy, heterocyclyl, substituted heterocyclyl,        heterocyclyloxy, substituted heterocyclyloxy, aminoacyl,        aminoacyloxy, carboxyl, carboxyl ester, carbonate ester, nitro        or halo;    -   each R⁴ independently is hydrogen, alkyl, substituted alkyl,        cycloalkyl, substituted cycloalkyl, heterocyclyl, substituted        heterocyclyl, carboxyl, carboxyl ester, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, acyl or M⁺ wherein M⁺, a        counterion, is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄, wherein each R⁹        independently is hydrogen or alkyl, and the nitrogen immediately        adjacent to R⁵ is anionic; or        -   R⁴ and R⁵ together with the intervening atom or atoms bound            thereto, form a heterocyclyl or substituted heterocyclyl            group;    -   each R⁵ independently is hydrogen, alkyl, substituted alkyl,        amino, cycloalkyl, substituted cycloalkyl, heterocyclyl,        substituted heterocyclyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, carboxyl, carboxyl ester or acyl;    -   R⁶ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl and        acyl;    -   R⁶ is a straight or branched chain C₁₋₆ alkylene group,        cycloalkylene or substituted cycloalkylene, linking the nitrogen        bearing R⁶ and the ring bearing Y    -   R⁷ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl;    -   R⁸ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl;    -   optionally, R⁷ and R⁸, together with nitrogen to which they are        bound, form a heterocyclyl or substituted heterocyclyl group;    -   wherein at least one of R⁷ and R⁸ is non-hydrogen; and    -   each of R⁶, R⁷, and R⁸ optionally are M⁺ wherein M⁺, a        counter-ion, is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄, wherein each R⁹        independently is hydrogen or alkyl, and the nitrogen bearing M⁺        is anionic.

In certain implementations, the invention provides compounds of formulaII:

wherein

-   -   R¹⁰ is a straight or branched chain C₁₋₆ alkylene group,        cycloalkylene or substituted cycloalkylene, and the remaining        variables are as defined as for formula I except that included        are compounds where R⁷ and R⁸ are simultaneously hydrogen.

Yet other implementations of the invention provide compounds of formulaIII:

wherein:

-   -   X is alkyl, substituted alkyl, hydroxy, alkoxy, substituted        alkoxy, amino, substituted amino, carboxyl, carboxyl ester,        —C(O)N(R⁴)R⁵, cyano, halo, nitro, alkenyl, substituted alkenyl,        alkynyl, substituted alkynyl, cycloalkynyl or substituted        cycloalkynyl; and the remaining variables are as defined for        compounds of formula I.

Another implementation of the invention provides compounds of formulaIV:

wherein:

-   -   X is alkyl, substituted alkyl, hydroxy, alkoxy, substituted        alkoxy, amino, substituted amino, carboxyl, carboxyl ester,        —C(O)N(R⁴)R⁵, cyano, halo, nitro, alkenyl, substituted alkenyl,        alkynyl, substituted alkynyl, cycloalkynyl or substituted        cycloalkynyl; and the remaining variables are as defined for        compounds of formula I.

Another embodiment is a method of inhibiting an activity of a JAKkinase, including contacting the JAK kinase with an amount of a compoundeffective to inhibit an activity of the JAK kinase where the compound isaccording to formula I as described herein. In one embodiment thecontact is made in vitro, in another embodiment the contact is made invivo.

Another embodiment is a method of treating a T-cell mediated autoimmunedisease, including administering to a patient suffering from such anautoimmune disease an amount of a compound effective to treat theautoimmune disease where the compound is according to formula I asdescribed herein.

Another embodiment is a method of treating or preventing allografttransplant rejection in a transplant recipient, including administeringto the transplant recipient an amount of a compound effective to treator prevent the rejection where the compound is according to formula I asdescribed herein. Administration in this context may include contactinga transplant organ with a compound or pharmaceutical compositiondescribed herein prior to transplant and/or concurrent withadministration to the transplant recipient.

Yet another embodiment is a method of treating or preventing a Type IVhypersensitivity reaction, including administering to a subject anamount of a compound of effective to treat or prevent thehypersensitivity reaction where the compound is according to formula Ias described herein.

Another embodiment is a method of treating or preventing an oculardisease or disorder, including administering to a subject an amount of acompound of effective to treat or prevent the ocular disease or disorderwhere the compound is according to formula I as described herein.

Another embodiment is a method of inhibiting a signal transductioncascade in which JAK3 kinase plays a role, including contacting a cellexpressing a receptor involved in such a signaling cascade with acompound where the compound is according to formula I as describedherein.

Another embodiment is a method of treating or preventing a JAKkinase-mediated disease, including administering to a subject an amountof compound effective to treat or prevent the JAK kinase-mediateddisease where the compound is according to formula I as describedherein.

Another embodiment is a pharmaceutical formulation including a compoundof formula I as described herein. Therapy using the2,4-pyrimidinediamine compounds and pharmaceutical formulationsdescribed herein can be applied alone, or it can be applied incombination with or adjunctive to other common immunosuppressivetherapies

Other embodiments include methods of using the compounds for screeningfor other agents used to treat or prevent a JAK kinase mediated disease.

More detailed description for these and other embodiments is providedbelow.

III. DETAILED DESCRIPTION A. Overview

The invention encompasses compounds having formula I and thecompositions and methods using these compounds in the treatment ofconditions in which modulation of the JAK pathway or inhibition of JAKkinases, particularly JAK3, are therapeutically useful.

B. Definitions

As used herein, the following definitions shall apply unless otherwiseindicated.

“Alkyl” refers to monovalent saturated aliphatic hydrocarbyl groupshaving from 1 to 10 carbon atoms and preferably 1 to 6 carbon atoms.This term includes, by way of example, linear and branched hydrocarbylgroups such as methyl (CH₃—), ethyl (CH₃CH₂—), n-propyl (CH₃CH₂CH₂—),isopropyl ((CH₃)₂CH—), n-butyl (CH₃CH₂CH₂CH₂—), isobutyl ((CH₃)₂CHCH₂—),sec-butyl ((CH₃)(CH₃CH₂)CH—), t-butyl ((CH₃)₃C—), n-pentyl(CH₃CH₂CH₂CH₂CH₂—), and neopentyl ((CH₃)₃CCH₂—).

“Alkylene” refers to divalent saturated aliphatic hydrocarbyl groupspreferably having from 1 to 6 and more preferably 1 to 3 carbon atomsthat are either straight-chained or branched. This term is exemplifiedby groups such as methylene (—CH₂—), ethylene (—CH₂CH₂—), n-propylene(—CH₂CH₂CH₂—), iso-propylene (—CH₂CH(CH₃)—) or (—CH(CH₃)CH₂—), and thelike.

“Substituted alkylene” refers to an alkylene group having from 1 to 3hydrogens replaced with substituents as described for carbons in thedefinition of “substituted” below.

“Alkoxy” refers to the group —O-alkyl, wherein alkyl is as definedherein. Alkoxy includes, by way of example, methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, t-butoxy, sec-butoxy, n-pentoxy, and the like.

“Acyl” refers to the groups H—C(O)—, alkyl-C(O)—, substitutedalkyl-C(O)—, alkenyl-C(O)—, substituted alkenyl-C(O)—, alkynyl-C(O)—,substituted alkynyl-C(O)—, cycloalkyl-C(O)—, substitutedcycloalkyl-C(O)—, cycloalkenyl-C(O)—, substituted cycloalkenyl-C(O)—,aryl-C(O)—, substituted aryl-C(O)—, heteroaryl-C(O)—, substitutedheteroaryl-C(O)—, heterocyclyl-C(O)—, and substitutedheterocyclyl-C(O)—, wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl, andsubstituted heterocyclyl are as defined herein. Acyl includes the“acetyl” group CH₃C(O)—.

“Acylamino” refers to the groups NR²⁰C(O)alkyl, —NR²⁰C(O)substitutedalkyl, N R²⁰C(O)cycloalkyl, —NR²⁰C(O)substituted cycloalkyl,—NR²⁰C(O)cycloalkenyl, —NR²⁰C(O)substituted cycloalkenyl,—NR²⁰C(O)alkenyl, —NR²⁰C(O)substituted alkenyl, —NR²⁰C(O)alkynyl,—NR²⁰C(O)substituted alkynyl, —NR²⁰C(O)aryl, —NR²⁰C(O)substituted aryl,—NR²⁰C(O)heteroaryl, —NR²⁰C(O)substituted heteroaryl,—NR²⁰C(O)heterocyclyl, and —NR²⁰C(O)substituted heterocyclyl, whereinR²⁰ is hydrogen or alkyl and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl, andsubstituted heterocyclyl are as defined herein.

“Acyloxy” refers to the groups alkyl-C(O)O—, substituted alkyl-C(O)O—,alkenyl-C(O)O—, substituted alkenyl-C(O)O—, alkynyl-C(O)O—, substitutedalkynyl-C(O)O—, aryl-C(O)O—, substituted aryl-C(O)O—, cycloalkyl-C(O)O—,substituted cycloalkyl-C(O)O—, cycloalkenyl-C(O)O—, substitutedcycloalkenyl-C(O)O—, heteroaryl-C(O)O—, substituted heteroaryl-C(O)O—,heterocyclyl-C(O)O—, and substituted heterocyclyl-C(O)O—, wherein alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclyl, and substituted heterocyclyl are as definedherein.

“Amino” refers to the group —NH₂.

“Aminocarbonyl” refers to the group —C(O)NR²¹R²², wherein R²¹ and R²²independently are selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclyl, and substituted heterocyclyl and where R²¹ andR²² are optionally joined together with the nitrogen bound thereto toform a heterocyclyl or substituted heterocyclyl group, and whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclyl, and substituted heterocyclyl areas defined herein.

“Aminothiocarbonyl” refers to the group —C(S)NR²¹R²², wherein R²¹ andR²² independently are selected from the group consisting of hydrogen,alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, aryl, substituted aryl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl,substituted heteroaryl, heterocyclyl, and substituted heterocyclyl andwhere R²¹ and R²² are optionally joined together with the nitrogen boundthereto to form a heterocyclyl or substituted heterocyclyl group, andwherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, awl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclyl, and substituted heterocyclyl are as definedherein.

“Aminocarbonylamino” refers to the group —NR²⁰C(O)NR²¹R²², wherein R²⁰is hydrogen or alkyl and R²¹ and R²² independently are selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, substitutedaryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclyl, andsubstituted heterocyclyl and where R²¹ and R²² are optionally joinedtogether with the nitrogen bound thereto to form a heterocyclyl orsubstituted heterocyclyl group, and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl andsubstituted heterocyclyl are as defined herein.

“Aminothiocarbonylamino” refers to the group —NR²⁰C(S)NR²¹R²², whereinR²⁰ is hydrogen or alkyl and R²¹ and R²² independently are selected fromthe group consisting of hydrogen, alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, substitutedawl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclyl, andsubstituted heterocyclyl and where R²¹ and R²² are optionally joinedtogether with the nitrogen bound thereto to form a heterocyclyl orsubstituted heterocyclyl group, and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl andsubstituted heterocyclyl are as defined herein.

“Aminocarbonyloxy” refers to the group —O—C(O)NR²¹R²², wherein R²¹ andR²² independently are selected from the group consisting of hydrogen,alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, aryl, substituted awl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl,substituted heteroaryl, heterocyclyl, and substituted heterocyclyl andwhere R²¹ and R²² are optionally joined together with the nitrogen boundthereto to form a heterocyclyl or substituted heterocyclyl group, andwherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, awl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclyl and substituted heterocyclyl are as definedherein.

“Aminosulfonyl” refers to the group —SO₂NR²¹R²², wherein R²¹ and R²²independently are selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclyl, substituted heterocyclyl and where R²¹ and R²²are optionally joined together with the nitrogen bound thereto to form aheterocyclyl or substituted heterocyclyl group and alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclyl and substituted heterocyclyl are as definedherein.

“Aminosulfonyloxy” refers to the group —O—SO₂NR²¹R²², wherein R²¹ andR²² independently are selected from the group consisting of hydrogen,alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, aryl, substituted aryl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl,substituted heteroaryl, heterocyclyl, and substituted heterocyclyl; R²¹and R²² are optionally joined together with the nitrogen bound theretoto form a heterocyclyl or substituted heterocyclyl group; and alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclyl, and substituted heterocyclyl are as definedherein.

“Aminosulfonylamino” refers to the group —NR²⁰—SO₂NR²¹R²², wherein R²⁰is hydrogen or alkyl and R²¹ and R²² independently are selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, substitutedaryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclyl, andsubstituted heterocyclyl and where R²¹ and R²² are optionally joinedtogether with the nitrogen bound thereto to form a heterocyclyl orsubstituted heterocyclyl group, and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, awl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl andsubstituted heterocyclyl are as defined herein.

“Sulfonylamino” refers to group —NR²¹SO₂R²², wherein R²¹ and R²²independently are selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclyl, and substituted heterocyclyl and where R²¹ andR²² are optionally joined together with the atoms bound thereto to forma heterocyclyl or substituted heterocyclyl group, and wherein alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclyl, and substituted heterocyclyl are as definedherein.

“Amidino” refers to the group —C(═NR³⁰)NR³¹R³², wherein R³¹ and R³²independently are selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclyl, and substituted heterocyclyl and where R³¹ andR³² are optionally joined together with the nitrogen bound thereto toform a heterocyclyl or substituted heterocyclyl group. R³⁰ is selectedfrom the group consisting of hydrogen, alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkynyl, substituted cycloalkynyl, aryl, substituted aryl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclyl,substituted heterocyclyl, nitro, nitroso, hydroxy, alkoxy, cyano,—N═N—N-alkyl, —N═N—N-substituted alkyl, —N(alkyl)SO₂-alkyl,—N(alkyl)SO₂-substituted alkyl, —N═N═N-alkyl, —N═N═N— substituted alkyl,acyl, —SO₂-alkyl and —SO₂-substituted alkyl, wherein alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkynyl, substituted cycloalkynyl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclyl, substitutedheterocyclyl, nitro, nitroso, hydroxy, alkoxy, and cyano are as definedherein. One of R³¹ and R³² along with R³⁰ are optionally joined togetherwith the nitrogens bound thereto and the intervening carbon of theguanidine group to form a cyclic amidine.

“Aryl” or “Ar” refers to a monovalent aromatic carbocyclic group of from6 to 14 carbon atoms having a single ring (e.g., phenyl) or multiplecondensed rings (e.g., naphthyl or anthryl) which condensed rings may ormay not be aromatic (e.g., 2-benzoxazolinone,2H-1,4-benzoxazin-3(4H)-one-7-yl, and the like), provided that the pointof attachment is through an atom of the aromatic aryl group. Preferredaryl groups include phenyl and naphthyl.

“Aryloxy” refers to the group —O-aryl, wherein awl is as defined herein,including, by way of example, phenoxy, naphthoxy, and the like.

“Arylthio” refers to the group —S-aryl, wherein aryl is as definedherein. In other embodiments, sulfur may be oxidized to —S(O)— or —SO₂—moieties. Depending on the pendant substitution, the sulfoxide mayimpart chirality to the molecule.

“Alkenyl” refers to straight chain or branched hydrocarbyl groups havingfrom 2 to 6 carbon atoms and preferably 2 to 4 carbon atoms and havingat least 1 and preferably from 1 to 2 sites of double bond unsaturation.Such groups are exemplified, for example, bi-vinyl, allyl, andbut-3-en-1-yl. Included within this term are the cis and trans isomersor mixtures of these isomers.

“Alkynyl” refers to straight or branched monovalent hydrocarbyl groupshaving from 2 to 6 carbon atoms and preferably 2 to 3 carbon atoms andhaving at least 1 and preferably from 1 to 2 sites of triple bondunsaturation. Examples of such alkynyl groups include acetylenyl(—C≡CH), and propargyl (—CH₂CCH).

“Alkynyloxy” refers to the group —O-alkynyl, wherein alkynyl is asdefined herein. Alkynyloxy includes, by way of example, ethynyloxy,propynyloxy, and the like.

“Carboxyl,” “carboxy” or “carboxylate” refers to —CO₂H or salts thereof.

“Carboxyl ester” or “carboxy ester” refers to the groups —C(O)O-alkyl,—C(O)O-substituted alkyl, —C(O)O-alkenyl, —C(O)O-substituted alkenyl,—C(O)O-alkynyl, C(O)O-substituted alkynyl, —C(O)O-aryl,—C(O)O-substituted aryl, —C(O)O-cycloalkyl, —C(O)O-substitutedcycloalkyl, —C(O)O-cycloalkenyl, —C(O)O-substituted cycloalkenyl,—C(O)O-heteroaryl, —C(O)O-substituted heteroaryl, —C(O)O-heterocyclyl,and —C(O)O-substituted heterocyclyl, wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted awl, heteroaryl, substituted heteroaryl, heterocyclyl, andsubstituted heterocyclyl are as defined herein.

“(Carboxyl ester)amino” refers to the groups —NR—C(O)O-alkyl,—NR—C(O)O-substituted alkyl, —NR—C(O)O-alkenyl, —NR—C(O)O-substitutedalkenyl, —NR—C(O)O-alkynyl, —NR—C(O)O-substituted alkynyl,—NR—C(O)O-aryl, —NR—C(O)O-substituted aryl, —NR—C(O)O-cycloalkyl,—NR—C(O)O-substituted cycloalkyl, —NR—C(O)O-cycloalkenyl,—NR—C(O)O-substituted cycloalkenyl, —NR—C(O)O-heteroaryl,—NR—C(O)O-substituted heteroaryl, —NR—C(O)O-heterocyclyl, and—NR—C(O)O-substituted heterocyclyl, wherein R is alkyl or hydrogen andalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclyl, and substituted heterocyclyl areas defined herein.

“(Carboxyl ester)oxy” or “carbonate” refers to the groups—O—C(O)O-alkyl, —O—C(O)O-substituted alkyl, —O—C(O)O-alkenyl,—O—C(O)O-substituted alkenyl, —O—C(O)O-alkynyl, —O—C(O)O-substitutedalkynyl, —O—C(O)O-aryl, —O—C(O)O-substituted aryl, —O—C(O)O-cycloalkyl,—O—C(O)O-substituted cycloalkyl, —O—C(O)O-cycloalkenyl,—O—C(O)O-substituted cycloalkenyl, —O—C(O)O-heteroaryl,—O—C(O)O-substituted heteroaryl, —O—C(O)O-heterocyclyl, and—O—C(O)O-substituted heterocyclyl, wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl, andsubstituted heterocyclyl are as defined herein.

“Cyano” or “nitrile” refers to the group —CN.

“Cycloalkyl” refers to cyclic alkyl groups of from 3 to 10 carbon atomshaving single or multiple cyclic rings including fused, bridged, andspiro ring systems. Examples of suitable cycloalkyl groups include, forinstance, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyland the like.

“Cycloalkenyl” refers to non-aromatic cyclic alkyl groups of from 3 to10 carbon atoms having single or multiple rings and having at least onedouble bond and preferably from 1 to 2 double bonds.

“Cycloalkynyl” refers to non-aromatic cycloalkyl groups of from 5 to 10carbon atoms having single or multiple rings and having at least onetriple bond.

“Cycloalkylene” refers to divalent cycloalkyl groups, wherein cycloalkylis as defined herein.

“Cycloalkoxy” refers to —O-cycloalkyl.

“Cycloalkylthio” refers to —S-cycloalkyl. In other embodiments, sulfurmay be oxidized to —S(O)— or —SO₂— moieties. Depending on the pendantsubstitution, the sulfoxide may impart chirality to the molecule.

“Cycloalkenyloxy” refers to ±0-cycloalkenyl.

“Cycloalkenylthio” refers to —S-cycloalkenyl. In other embodiments,sulfur may be oxidized to sulfinyl or sulfonyl moieties. Depending onthe pendant substitution, the sulfoxide may impart chirality to themolecule.

“Guanidino” refers to the group —NHC(═NH)NH₂.

“Substituted guanidino” refers to the group —NR³³C(═NR³³)N(R³³)₂,wherein each R³³ independently is selected from the group consisting ofhydrogen, alkyl, substituted alkyl, aryl, substituted awl, heteroaryl,substituted heteroaryl, heterocyclyl, and substituted heterocyclyl; twoR groups attached to a common guanidino nitrogen atom are optionallyjoined together with the nitrogen bound thereto to form a heterocyclylor substituted heterocyclyl group, provided that at least one R is nothydrogen; and said substituents are as defined herein. Two R³³ groups ondistinct nitrogens are optionally joined together with the nitrogensbound thereto and the intervening carbon of the guanidine group to forma cyclic guanidine.

“Halo” or “halogen” refers to fluoro, chloro, bromo, and iodo and ispreferably fluoro or chloro.

“Hydroxy” or “hydroxyl” refers to the group —OH.

“Heteroaryloxy” refers to —O-heteroaryl.

“Heteroarylthio” refers to the group —S-heteroaryl. In otherembodiments, sulfur may be oxidized to —S(O)— or —SO₂— moieties. Thesulfoxide may exist as one or more stereoisomers.

“Heterocyclyl” in the broadest sense includes aromatic and non-aromaticring systems and more specifically refers to a stable three- tofifteen-membered ring radical that consists of carbon atoms and from oneto five heteroatoms. For purposes of this invention, the heterocyclylradical can be a monocyclic, bicyclic or tricyclic ring system, whichcan include fused or bridged ring systems as well as spirocyclicsystems; and the nitrogen, phosphorus, carbon or sulfur atoms in theheterocyclyl radical can be optionally oxidized to various oxidationstates. In a specific example, the group —S(O)₀₋₂—, refers to—S-(sulfide), —S(O)— (sulfoxide), and —SO₂— (sulfone) linkages. Forconvenience, nitrogens, particularly but not exclusively, those definedas annular aromatic nitrogens, are meant to include their correspondingN-oxide form, although not explicitly defined as such in a particularexample. Thus, for a compound having, for example, a pyridyl ring; thecorresponding pyridyl-N-oxide is meant to be included as anothercompound of the invention. In addition, annular nitrogen atoms can beoptionally quaternized. “Heterocycle” includes heteroaryl andheteroalicyclyl, that is a heterocyclyl ring can be partially or fullysaturated or aromatic. Thus a term such as “heterocyclylalkyl” includesheteroalicyclylalkyls and heteroarylalkyls. Examples of heterocyclylradicals include, but are not limited to, azetidinyl, acridinyl,benzodioxolyl, benzodioxanyl, benzofuranyl, carbazoyl, cinnolinyl,dioxolanyl, indolizinyl, naphthyridinyl, perhydroazepinyl, phenazinyl,phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl,quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrazoyl,tetrahydroisoquinolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl,4-piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl,imidazolinyl, imidazolidinyl, dihydropyridinyl, tetrahydropyridinyl,pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl,oxazolidinyl, triazolyl, isoxazolyl, isoxazolidinyl, morpholinyl,thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl, quinuclidinyl,isothiazolidinyl, indolyl, isoindolyl, indolinyl, isoindolinyl,octahydroindolyl, octahydroisoindolyl, quinolyl, isoquinolyl,decahydroisoquinolyl, benzimidazolyl, thiadiazolyl, benzopyranyl,benzothiazolyl, benzoxazolyl, furyl, diazabicycloheptane, diazapane,diazepine, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothieliyl,thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone,dioxaphospholanyl, and oxadiazolyl.

“Heteroaryl” refers to an aromatic group of having from 1 to 10 annularcarbon atoms and 1 to 4 annular heteroatoms within the ring. Heteroarylgroups have at least one aromatic ring component, but heteroaryls can befully unsaturated or partially unsaturated. If any aromatic ring in thegroup has a heteroatom, then the group is a heteroaryl, even, forexample, if other aromatic rings in the group have no heteroatoms. Forexample, 2H-pyrido[3,2-b][1,4]oxazin-3(4H)-one-7-yl, indolyl andbenzimidazolyl are “heteroaryls.” Heteroaryl groups can have a singlering (e.g., pyridinyl, imidazolyl or furyl) or multiple condensed rings(e.g., indolizinyl, quinolinyl, benzimidazolyl or benzothienyl), wherethe condensed rings may or may not be aromatic and/or contain aheteroatom, provided that the point of attachment to the parent moleculeis through an atom of the aromatic portion of the heteroaryl group. Inone embodiment, the nitrogen and/or sulfur ring atom(s) of theheteroaryl group are optionally oxidized to provide for the N-oxide(N→O), sulfinyl, or sulfonyl moieties. Compounds described hereincontaining phosphorous, in a heterocyclyl ring or not, include theoxidized forms of phosphorous. Heteroaryl groups are monocyclic,bicyclic, tricyclic or tetracyclic, and the like.

“Heteroaryloxy” refers to —O-heteroaryl.

“Heteroarylene” generically refers to any heteroaryl that has at leasttwo groups attached thereto. For a more specific example, “pyridylene”refers to a divalent pyridyl ring radical. A pyridylene, thus can havemore than two groups attached, but is defined by a minimum of twonon-hydrogen groups attached thereto.

“Heteroalicyclic” refers specifically to a non-aromatic heterocyclylradical. A heteroalicyclic may contain unsaturation, but is notaromatic. As mentioned, aryls and heteroaryls are attached to the parentstructure via an aromatic ring. So, e.g.,2H-1,4-benzoxazin-3(4H)-one-4-yl is a heteroalicyclic, while2H-1,4-benzoxazin-3(4H)-one-7-yl is an aryl. In another example,2H-pyrido[3,2-b][1,4]oxazin-3(4H)-one-4-yl is a heteroalicyclic, while2H-pyrido[3,2-b][1,4]oxazin-3(4H)-one-7-yl is a heteroaryl.

“Heterocyclylalkyl” refers to a heterocyclyl group linked to the parentstructure via e.g an alkylene linker, for example(tetrahydrofuran-3-yl)methyl- or (pyridin-4-yl)methyl

“Heterocyclyloxy” refers to the group —O-heterocycyl.

“Heterocyclylthio” refers to the group —S-heterocycyl. In otherembodiments, sulfur may be oxidized to —S(O)— or SO₂— moieties. Thesulfoxide may exist as one or more stereoisomers.

“Nitro” refers to the group —NO₂.

“Nitroso” refers to the group —NO.

“Oxo” refers to the atom (═O).

“Sulfonyl” refers to the group SO₂-alkyl, SO₂-substituted alkyl,SO₂-alkenyl, SO₂-substituted alkenyl, SO₂-hcycloalkyl, SO₂-substitutedcylcoalkyl, SO₂-cycloalkenyl, SO₂-substituted cylcoalkenyl, SO₂-aryl,SO₂-substituted aryl, SO₂-heteroaryl, SO₂-substituted heteroaryl,SO₂-heterocyclyl, and SO₂-substituted heterocyclyl, wherein alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclyl, and substituted heterocyclyl are as definedherein. Sulfonyl includes groups such as methyl-SO₂—, phenyl-SO₂—, and4-methylphenyl-SO₂—.

“Sulfonyloxy” refers to the group OSO₂-alkyl, OSO₂-substituted alkyl,OSO₂-alkenyl, OSO₂-substituted alkenyl, OSO₂-cycloalkyl,OSO₂-substituted cylcoalkyl, OSO₂-cycloalkenyl, OSO₂-substitutedcylcoalkenyl, OSO₂-aryl, OSO₂-substituted aryl, OSO₂-heteroaryl,OSO₂-substituted heteroaryl, OSO₂-heterocyclyl, and OSO₂ substitutedheterocyclyl, wherein alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclyl, and substitutedheterocyclyl are as defined herein.

“Thioacyl” refers to the groups H—C(S)—, alkyl-C(S)—, substitutedalkyl-C(S)—, alkenyl-C(S)—, substituted alkenyl-C(S)—, alkynyl-C(S)—,substituted alkynyl-C(S)—, cycloalkyl-C(S)—, substitutedcycloalkyl-C(S)—, cycloalkenyl-C(S)—, substituted cycloalkenyl-C(S)—,aryl-C(S)—, substituted aryl-C(S)—, heteroaryl-C(S)—, substitutedheteroaryl-C(S)—, heterocyclyl-C(S)—, and substitutedheterocyclyl-C(S)—, wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclyl, andsubstituted heterocyclyl are as defined herein.

“Thiol” refers to the group —SH.

“Thioxo” refers to the atom (═S).

“Alkylthio” refers to the group —S-alkyl, wherein alkyl is as definedherein. In other embodiments, sulfur may be oxidized to —S(O)—. Thesulfoxide may exist as one or more stereoisomers.

Unless indicated otherwise, the nomenclature of substituents that arenot explicitly defined herein are arrived at by naming the terminalportion of the functionality followed by the adjacent functionalitytoward the point of attachment. For example, the substituent“arylalkyloxycarbonyl” refers to the group (aryl)-(alkyl)-O—C(O)—.

The term “substituted,” when used to modify a specified group orradical, means that one or more hydrogen atoms of the specified group orradical are each, independently of one another, replaced with the sameor different substituent groups as defined below.

Substituent groups for substituting for one or more hydrogens (any twohydrogens on a single carbon can be replaced with ═O, ═NR⁷⁰, ═N—OR⁷⁰,═N₂ or ═S) on saturated carbon atoms in the specified group or radicalare, unless otherwise specified, —R⁶⁰, halo, ═O, —OR⁷⁰, —SR⁷⁰, —NR⁸⁰R⁸⁰,trihalomethyl, —CN, —OCN, —SCN, —NO, —NO₂, ═N₂, —N₃, —SO₂R⁷⁰, —SO₂O⁻M⁺,—SO₂OR⁷⁰, —OSO₂R⁷⁰, —OSO₂O⁻M⁺, —OSO₂OR⁷⁰, —P(O)(O⁻)₂(M⁺)₂,—P(O)(OR⁷⁰)O⁻M⁺, —P(O)(OR⁷⁰)₂, —C(O)R⁷⁰, —C(S)R⁷⁰, —C(NR⁷⁰)R⁷⁰,—C(O)O⁻M⁺, —C(O)OR⁷⁰, —C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰, —C(NR⁷⁰)NR⁸⁰R₈₀,—OC(O)R⁷⁰, —OC(S)R⁷⁰, —OC(O) O⁻M⁺, —OC(O)OR⁷⁰, —OC(S)OR⁷⁰, —NR⁷⁰C(O)R⁷⁰,—NR⁷⁰C(S)R⁷⁰, —NR⁷⁰CO₂ ⁻M⁺, —NR⁷⁰CO₂R⁷⁰, —NR⁷⁰C(S)OR⁷⁰,—NR⁷⁰C(O)NR⁸⁰R⁸⁰, NR⁷⁰C(NR⁷⁰)R⁷⁰ and —NR⁷⁰C(NR⁷⁰)NR⁸⁰R⁸⁰, where R⁶⁰ isselected from the group consisting of optionally substituted alkyl,cycloalkyl, heteroalkyl, heterocycloalkylalkyl, cycloalkylalkyl, aryl,arylalkyl, heteroaryl and heteroarylalkyl, each R⁷⁰ is independentlyhydrogen or R⁶⁰; each R⁸⁰ is independently R⁷⁰ or alternatively, twoR^(80′)s, taken together with the nitrogen atom to which they arebonded, form a 5-, 6- or 7-membered heterocycloalkyl which mayoptionally include from 1 to 4 of the same or different additionalheteroatoms selected from the group consisting of O, N and S, of which Nmay have —H or C₁-C₃ alkyl substitution; and each M⁺ is a counter ionwith a net single positive charge. Each M⁺ may independently be, forexample, an alkali ion, such as K⁺, Na⁺, Li⁺; an ammonium ion, such as⁺N(R⁶⁰)₄; or an alkaline earth ion, such as [Ca²⁺]_(0.5), [Mg²⁺]_(0.5),or [Ba²⁺]_(0.5) (“subscript 0.5 means e.g. that one of the counter ionsfor such divalent alkali earth ions can be an ionized form of a compoundof the invention and the other a typical counter ion such as chloride,or two ionized compounds of the invention can serve as counter ions forsuch divalent alkali earth ions, or a doubly ionized compound of theinvention can serve as the counter ion for such divalent alkali earthions). As specific examples, NR⁸⁰R⁸⁰ is meant to include —NH₂,—NH-alkyl, N-pyrrolidinyl, N-piperazinyl, 4N-methyl-piperazin-1-yl andN-morpholinyl.

Substituent groups for hydrogens on unsaturated carbon atoms in“substituted” alkene, alkyne, aryl and heteroaryl groups are, unlessotherwise specified, —R⁶⁰, halo, —O⁻M⁺, —OR⁷⁰, —SR⁷⁰, —S⁻M⁺, —NR⁸⁰R⁸⁰,trihalomethyl, —CF₃, —CN, —OCN, —SCN, —NO, —NO₂, —N₃, —SO₂R⁷⁰, —SO₃ ⁻M⁺,—SO₃R⁷⁰, —OSO₂R⁷⁰, —OSO₃ ⁻M⁺, —OSO₃R⁷⁰, —PO₃ ⁻²(M⁺)₂, —P(O)(OR⁷⁰)O⁻M⁺,—P(O)(OR⁷⁰)₂, —C(O)R⁷⁰, —C(S)R⁷⁰, —C(NR⁷⁰)R⁷⁰, —CO₂ ⁻M⁺, —CO₂R⁷⁰,—C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰, —C(NR⁷⁰)NR⁸⁰R⁸⁰, —OC(O)R⁷⁰, —OC(S)R⁷⁰, —OCO₂⁻M⁺, —OCO₂R⁷⁰, —OC(S)OR⁷⁰, —NR⁷⁰C(O)R⁷⁰, —NR⁷⁰C(S)R⁷⁰, —NR⁷⁰CO₂ ⁻M⁺,—NR⁸⁰CO₂R⁷⁰, —NR⁷⁰C(S)OR⁷⁰, —NR⁷⁰C(O)NR⁸⁰R⁸⁰, —NR⁷⁰C(NR⁷⁰)R⁷⁰ and—NR⁷⁰C(NR⁷⁰)NR⁸⁰R⁸⁰, where R⁶⁰, R⁷⁰, R⁸⁰ and M⁺ are as previouslydefined, provided that in case of substituted alkene or alkyne, thesubstituents are not —O⁻M⁺, —OR⁷⁰, —SR⁷⁰, or —S⁻M⁻.

Substituent groups for hydrogens on nitrogen atoms in “substituted”heteroalkyl and cycloheteroalkyl groups are, unless otherwise specified,—R⁶⁰, —O⁻M⁺, —OR⁷⁰, —SR⁷⁰, —S⁻M⁺, —NR⁸⁰R⁸⁰, trihalomethyl, —CF₃, —CN,—NO, —NO₂, —S(O)₂R⁷⁰, —S(O)₂O⁻M⁺, —S(O)₂OR⁷⁰, —OS(O)₂R⁷⁰, —OS(O)₂O⁻M⁺,—OS(O)₂OR⁷⁰, —P(O)(O⁻)₂(M⁺)₂, —P(O)(OR⁷⁰)O⁻M⁺, —P(O)(OR⁷⁰)(OR⁷⁰),—C(O)R⁷⁰, —C(S)R⁷⁰, —C(NR⁷⁰)R⁷⁰, —C(O)OR⁷⁰, —C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰,—C(NR⁷⁰)NR⁸⁰R⁸⁰, —OC(O)R⁷⁰, —OC(S)R⁷⁰, —OC(O)OR⁷⁰, —OC(S)OR⁷⁰,—NR⁷⁰C(O)R⁷⁰, —NR⁷⁰C(S)R⁷⁰, —NR⁷⁰C(O)OR⁷⁰, —NR⁷⁰C(S)OR⁷⁰,—NR⁷⁰C(O)NR⁸⁰R⁸⁰, —NR⁷⁰C(NR⁷⁰)R⁷⁰ and —NR⁸⁰C(NR⁷⁰)NR⁸⁰R⁸⁰, where R⁶⁰,R⁷⁰, R⁸⁰ and M⁺ are as previously defined.

In a preferred embodiment, a group that is substituted has 1, 2, 3, or 4substituents, 1, 2, or 3 substituents, 1 or 2 substituents, or 1substituent.

It is understood that in all substituted groups defined above, polymersarrived at by defining substituents with further substituents tothemselves (e.g., substituted aryl having a substituted awl group as asubstituent which is itself substituted with a substituted awl group,which is further substituted by a substituted aryl group, etc.) are notintended for inclusion herein. In such cases, the maximum number of suchsubstitutions is three. For example, serial substitutions of substitutedaryl groups are limited to substituted aryl-(substitutedaryl)-substituted awl.

“Patient” refers to human and non-human animals, especially mammals.

“Pharmaceutically acceptable salt” refers to pharmaceutically acceptablesalts of a compound, which salts are derived from a variety of organicand inorganic counter ions well known in the art and include, by way ofexample only, sodium, potassium, calcium, magnesium, ammonium,tetraalkylammonium, and the like; and when the molecule contains a basicfunctionality, salts of organic or inorganic acids, such ashydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate,oxalate, and the like.

“Prodrug” refers to a derivative of an active 4-pyrimidineamine compound(drug) that may require a transformation under the conditions of use,such as within the body, to release the active 2,4-pyrimidinediaminedrug. Prodrugs are frequently, but not necessarily, pharmacologicallyinactive until converted into the active drug. Prodrugs are typicallyobtained by masking one or more functional groups in an active2,4-pyrimidinediamine drug believed to be in part required for activitywith a progroup (defined below) to form a promoiety which undergoes atransformation, such as cleavage, under the specified conditions of useto release the functional group, and hence the active2,4-pyrimidinediamine drug. The cleavage of the promoiety may proceedspontaneously, such as by way of a hydrolysis reaction, or it can becatalyzed or induced by another agent, such as an enzyme, light, an acidor base, or a change of or exposure to a physical or environmentalparameter, such as temperature. The agent can be endogenous to theconditions of use, such as an enzyme present in the cells to which theprodrug is administered or the acidic conditions of the stomach, or itcan be supplied exogenously.

“Progroup” refers to a type of protecting group that, when used to maska functional group within an active 2,4-pyrimidinediamine drug to form apromoiety, converts the drug into a prodrug. Progroups are typicallyattached to the functional group of the drug via bonds that arecleavable under specified conditions of use. Thus, a progroup is thatportion of a promoiety that cleaves to release the functional groupunder the specified conditions of use. As a specific example, an amidepromoiety of the formula —NH—C(O)CH3 includes the progroup C(O)CH3.

“Pharmaceutically effective amount” and “therapeutically effectiveamount” refer to an amount of a compound sufficient to treat a specifieddisorder or disease or one or more of its symptoms and/or to prevent theoccurrence of the disease or disorder. In reference to tumorigenicproliferative disorders, a pharmaceutically or therapeutically effectiveamount includes an amount sufficient to, among other things, cause thetumor to shrink or decrease the growth rate of the tumor.

“Solvate” refers to a complex formed by combination of solvent moleculeswith molecules or ions of the solute. The solvent can be an organiccompound, an inorganic compound, or a mixture of both. Some examples ofsolvents include, but are not limited to, methanol,N,N-dimethylformamide, tetrahydrofuran, dimethylsulfoxide, and water.

One of ordinary skill in the art will appreciate that compounds of theinvention may exhibit the phenomena of tautomerism, conformationalisomerism, geometric isomerism, and/or optical isomerism. For example,the compounds and prodrugs of the invention can include one or morechiral centers and/or double bonds and as a consequence can exist asstereoisomers, such as double-bond isomers (i.e., geometric isomers),enantiomers, diasteromers, and mixtures thereof, such as racemicmixtures. As another example, the compounds of the invention can existin several tautomeric forms, including the enol form, the keto form, andmixtures thereof. As the various compound names, formulae and compounddrawings within the specification and claims can represent only one ofthe possible tautomeric, conformational isomeric, optical isomeric, orgeometric isomeric forms, it would be understood that the inventionencompasses any tautomeric, conformational isomeric, optical isomeric,and/or geometric isomeric forms of the compounds described herein, aswell as mixtures of these various different isomeric forms. In cases oflimited rotation, e.g. around the 2,4-pryimidinediamine core structure,atropisomers are also possible and are also specifically included in thecompounds of the invention. It is intended that the compoundsencompassed herein are, with the exception of forms of isomerism,chemically stable and isolable.

As is understood by one of ordinary skill in the art, certain atomsoccur in more than one isotopic form. For example hydrogen occurs asprotium (¹H), deuterium (²H) and tritium (³H), and carbon occursnaturally as three different isotopes, ¹²C, ¹³C and ¹⁴C. Thus thepresently disclosed formulas include compounds having one or moredifferent isotopic forms of certain elements, including hydrogen andcarbon. In one embodiment of the disclosure, the presently disclosedcompounds are provided in isotopically enriched form. In particularexamples, compounds of formula I are enriched in deuterium relative toprotium.

Deuterium has a natural abundance of about 0.015%. Accordingly, forapproximately every 6,500 hydrogen atoms occurring in nature, there isone deuterium atom. Disclosed herein are compounds enriched in deuteriumat one or more positions. Thus, deuterium containing compounds of thedisclosure have deuterium at one or more positions (as the case may be)in an abundance of greater than 0.015%.

In one embodiment, a compound of formula (I), at a position designatedas having deuterium, has a minimum isotopic enrichment factor of atleast 2000 (30% deuterium incorporation) at each atom designated asdeuterium in the compound, or at least 3000 (45% deuteriumincorporation).

In other embodiments, a compound of formula (I) has an isotopicenrichment factor for each designated deuterium atom of at least 3500(52.5% deuterium incorporation at each designated deuterium atom), atleast 4000 (60% deuterium incorporation), at least 4500 (67.5% deuteriumincorporation), at least 5000 (75% deuterium incorporation), at least5500 (82.5% deuterium incorporation), at least 6000 (90% deuteriumincorporation), at least 6333.3 (95% deuterium incorporation), at least6466.7 (97% deuterium incorporation), at least 6600 (99% deuteriumincorporation), or at least 6633.3 (99.5% deuterium incorporation).

Similarly, it is understood that the above definitions are not intendedto include impermissible substitution patterns (e.g., methyl substitutedwith 5 fluoro groups). Such impermissible substitution patterns areeasily recognized by a person having ordinary skill in the art.

C. Compounds of the Invention

Disclosed herein are novel 2,4-pyrimidinediamine compounds, prodrugs ofthe compounds, methods of making the compounds, and methods of usingthese compounds in the treatment of conditions in which targeting of theJAK pathway or modulation, including inhibition, of JAK kinases,particularly JAK3, are therapeutically useful. These conditions include,but are not limited to, leukemia, lymphoma, transplant rejection (e.g.,pancreas islet transplant rejection, heart transplant rejection, kidneytransplant rejection, liver transplant rejection, lung transplantrejection), bone marrow transplant applications (e.g., graft-versus-hostdisease), autoimmune diseases (e.g., diabetes), and inflammation (e.g.,asthma, allergic reactions, ocular disorders). Given the severity andprevalence of these diseases and conditions, new therapies are needed.

Generally, the 2,4-pyrimidinediamine compounds of the invention arecharacterized by a bis-phenyl 2,4-pyrimidinediamine where one phenylgroup bears at least a sulfonamide and the other phenyl group bears atleast a sulfonyl urea. More specifically, a compound of formula I:

wherein:

-   -   p is 0, 1, 2, or 3;    -   q is 0, 1, 2, or 3;    -   X is hydrogen, alkyl, substituted alkyl, hydroxy, alkoxy,        substituted alkoxy, amino, substituted amino, carboxyl, carboxyl        ester, —C(O)N(R⁴)R⁵, cyano, halo, nitro, alkenyl, substituted        alkenyl, alkynyl, substituted alkynyl, cycloalkynyl or        substituted cycloalkynyl;    -   Y is a straight or branched chain C₁₋₆ alkylene group,        cycloalkylene or substituted cycloalkylene;    -   R¹ is hydrogen, alkyl, substituted alkyl, cycloalkyl or        substituted cycloalkyl;    -   each R² independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, amino, substituted amino, aryl, substituted        aryl, aryloxy, substituted aryloxy, cyano, cycloalkyl,        substituted cycloalkyl, cycloalkoxy, substituted cycloalkoxy,        heteroaryl, substituted heteroaryl, heteroaryloxy, substituted        heteroaryloxy, heterocyclyl, substituted heterocyclyl,        heterocyclyloxy, substituted heterocyclyloxy, aminoacyl,        aminoacyloxy, carboxyl, carboxyl ester, carbonate ester,        —C(O)N(R⁴)R⁵, nitro or halo;    -   each R³ independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, amino, substituted amino, aryl, substituted        aryl, aryloxy, substituted aryloxy, cyano, cycloalkyl,        substituted cycloalkyl, cycloalkoxy, substituted cycloalkoxy,        heteroaryl, substituted heteroaryl, heteroaryloxy, substituted        heteroaryloxy, heterocyclyl, substituted heterocyclyl,        heterocyclyloxy, substituted heterocyclyloxy, aminoacyl,        aminoacyloxy, carboxyl, carboxyl ester, carbonate ester, nitro        or halo;    -   each R⁴ independently is hydrogen, alkyl, substituted alkyl,        cycloalkyl, substituted cycloalkyl, heterocyclyl, substituted        heterocyclyl, carboxyl, carboxyl ester, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, acyl or M⁺, wherein M⁺, a        counterion, is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄, wherein each R⁹        independently is hydrogen or alkyl, and the nitrogen immediately        adjacent to R⁵ is anionic; or        -   R⁴ and R⁵ together with the intervening atom or atoms bound            thereto, form a heterocyclyl or substituted heterocyclyl            group;    -   each R⁵ independently is hydrogen, alkyl, substituted alkyl,        amino, cycloalkyl, substituted cycloalkyl, heterocyclyl,        substituted heterocyclyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, carboxyl, carboxyl ester or acyl;    -   R⁶ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl and        acyl;    -   R⁶ is a straight or branched chain C₁₋₆alkylene group,        cycloalkylene or substituted cycloalkylene, linking the nitrogen        bearing R⁶ and the ring bearing Y    -   R⁷ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl;    -   R⁸ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl;    -   optionally, R⁷ and R⁸, together with nitrogen to which they are        bound, form a heterocyclyl or substituted heterocyclyl group;    -   wherein at least one of R⁷ and R⁸ is non-hydrogen; and    -   each of R⁶, R⁷, and R⁸ optionally are M⁺, wherein M⁺ a        counter-ion, is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄, wherein each R⁹        independently is hydrogen or alkyl, and the nitrogen bearing M⁺        is anionic.

One embodiment is a compound of structural formula I where R¹ ishydrogen. In another embodiment, where R¹ is hydrogen, X is alkyl,substituted alkyl or halo.

Another embodiment is a compound of structural formula I where R⁴ ishydrogen and R⁵ is hydrogen.

Another embodiment is a compound of structural formula I where Y ismethylene.

Another embodiment is a compound of structural formula I where R⁶ ishydrogen, R⁷ is C₁₋₃ alkyl and R⁸ is C₁₋₃ alkyl.

Another embodiment is a compound of structural formula I where each ofR² and R³, independently, is lower alkyl or lower alkoxy.

Another embodiment is a compound of structural formula II:

wherein

-   -   p is 0, 1, 2, or 3;    -   q is 0, 1, 2, or 3;    -   X is hydrogen, alkyl, substituted alkyl, hydroxy, alkoxy,        substituted alkoxy, amino, substituted amino, carboxyl, carboxyl        ester, —C(O)N(R⁴)R⁵, cyano, halo, nitro, alkenyl, substituted        alkenyl, alkynyl, substituted alkynyl, cycloalkynyl or        substituted cycloalkynyl;    -   Y is a straight or branched chain C₁₋₆ alkylene group,        cycloalkylene or substituted cycloalkylene;    -   R¹ is hydrogen, alkyl, substituted alkyl, cycloalkyl or        substituted cycloalkyl;    -   each R² independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, amino, substituted amino, aryl, substituted        aryl, aryloxy, substituted aryloxy, cyano, cycloalkyl,        substituted cycloalkyl, cycloalkoxy, substituted cycloalkoxy,        heteroaryl, substituted heteroaryl, heteroaryloxy, substituted        heteroaryloxy, heterocyclyl, substituted heterocyclyl,        heterocyclyloxy, substituted heterocyclyloxy, aminoacyl,        aminoacyloxy, carboxyl, carboxyl ester, carbonate ester,        —C(O)N(R⁴)R⁵, nitro or halo;    -   each R³ independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, amino, substituted amino, aryl, substituted        aryl, aryloxy, substituted aryloxy, cyano, cycloalkyl,        substituted cycloalkyl, cycloalkoxy, substituted cycloalkoxy,        heteroaryl, substituted heteroaryl, heteroaryloxy, substituted        heteroaryloxy, heterocyclyl, substituted heterocyclyl,        heterocyclyloxy, substituted heterocyclyloxy, aminoacyl,        aminoacyloxy, carboxyl, carboxyl ester, carbonate ester, nitro        or halo;    -   each R⁴ independently is hydrogen, alkyl, substituted alkyl,        cycloalkyl, substituted cycloalkyl, heterocyclyl, substituted        heterocyclyl, carboxyl, carboxyl ester, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, acyl or M⁺ wherein M⁺ is K⁺,        Na⁺, Li⁺ or ⁺N(R⁹)₄, wherein each R⁹ independently is hydrogen        or alkyl, and the nitrogen immediately adjacent to R⁵ is        anionic; or        -   R⁴ and R⁵ together with the intervening atom or atoms bound            thereto, form a heterocyclyl or substituted heterocyclyl            group;    -   each R⁵ independently is hydrogen, alkyl, substituted alkyl,        amino, cycloalkyl, substituted cycloalkyl, heterocyclyl,        substituted heterocyclyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, carboxyl, carboxyl ester or acyl;    -   R⁷ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl;    -   R⁸ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl; or    -   R⁷ and R⁸ together, with nitrogen to which they are bound, form        a heterocyclyl or substituted heterocyclyl group;    -   R¹⁰ is a straight or branched chain C₁₋₆ alkylene group,        cycloalkylene or substituted cycloalkylene; and    -   each of R⁷ and R⁸ optionally are M⁺, wherein M⁺ is K⁺, Na⁺, Li⁺        or ⁺N(R⁹)₄, wherein each R⁹ independently is hydrogen or alkyl,        and the nitrogen bearing M⁺ is anionic.

Another embodiment is a compound of structural formula II, where p iszero and each of R⁶ and Y, independently, is a straight or branchedchain C₁₋₆ alkylene group.

Another embodiment is a compound of structural formula II where each ofR⁶ and Y, independently, is methylene or ethylene.

Another embodiment is a compound of structural formula III:

wherein:

-   -   p is 0, 1, 2, or 3;    -   q is 0, 1, 2, or 3;    -   X is alkyl, substituted alkyl, hydroxy, alkoxy, substituted        alkoxy, amino, substituted amino, carboxyl, carboxyl ester,        —C(O)N(R⁴)R⁵, cyano, halo, nitro, alkenyl, substituted alkenyl,        alkynyl, substituted alkynyl, cycloalkynyl or substituted        cycloalkynyl;    -   each R² independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, cycloalkyl, substituted cycloalkyl,        cycloalkoxy, substituted cycloalkoxy, heterocyclyl, substituted        heterocyclyl, heterocyclyloxy, substituted heterocyclyloxy,        —C(O)N(R⁴)R⁵ or halo;    -   each R³ independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, cycloalkyl, substituted cycloalkyl,        cycloalkoxy, substituted cycloalkoxy, heterocyclyl, substituted        heterocyclyl, heterocyclyloxy, substituted heterocyclyloxy or        halo;    -   R⁶ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl;    -   R⁷ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl;    -   R⁸ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl; or    -   R⁷ and R⁸ together, with nitrogen to which they are bound, form        a heterocyclyl or substituted heterocyclyl group;    -   wherein at least one of R⁷ and R⁸ is non-hydrogen; and    -   each of R⁶, R⁷, and R⁸ optionally are substituted with M⁺,        wherein M⁺ is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄, wherein each R⁹        independently is hydrogen or alkyl, and the nitrogen bearing M⁺        is anionic.

Another embodiment is a compound of structural formula IV:

wherein:

-   -   p is 0, 1, 2, or 3;    -   q is 0, 1, 2, or 3;    -   X is alkyl, substituted alkyl, hydroxy, alkoxy, substituted        alkoxy, amino, substituted amino, carboxyl, carboxyl ester,        —C(O)N(R⁴)R⁵, cyano, halo, nitro, alkenyl, substituted alkenyl,        alkynyl, substituted alkynyl, cycloalkynyl or substituted        cycloalkynyl;    -   each R² independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, cycloalkyl, substituted cycloalkyl,        cycloalkoxy, substituted cycloalkoxy, heterocyclyl, substituted        heterocyclyl, heterocyclyloxy, substituted heterocyclyloxy,        —C(O)N(R⁴)R⁵ or halo;    -   each R³ independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, cycloalkyl, substituted cycloalkyl,        cycloalkoxy, substituted cycloalkoxy, heterocyclyl, substituted        heterocyclyl, heterocyclyloxy, substituted heterocyclyloxy or        halo;    -   R⁶ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl;    -   R⁷ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl;    -   R⁸ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl; or    -   R⁷ and R⁸ together, with nitrogen to which they are bound, form        a heterocyclyl or substituted heterocyclyl group;    -   wherein at least one of R⁷ and R⁸ is non-hydrogen; and    -   each of R⁶, R⁷, and R⁸ optionally are substituted with M⁺        wherein M⁺ is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄, wherein each R⁹        independently is hydrogen or alkyl, and the nitrogen bearing M⁺        is anionic.

Another embodiment is a compound selected from Tables I and II, orstereoisomer, tautomer, prodrug, solvate, or pharmaceutically acceptablesalt thereof.

TABLE I I

# —YN(R⁶)SO₂N(R⁷)R⁸ R² R¹ X R³ —SO₂N(R⁴)R⁵ I-1  4-CH₂NHSO₂N(CH₃)₂ — Hchloro — 3-SO₂NH₂ I-2  4-CH₂NHSO₂N(CH₃)₂ — H chloro — 4-SO₂NH₂ I-3 4-CH₂NHSO₂N(CH₃)₂ — H methyl — 3-SO₂NH₂ I-4  4-CH₂NHSO₂N(CH₃)₂ — Hmethyl — 4-SO₂NH₂ I-5  4-CH₂NHSO₂NHCH₂CH₃ — H chloro — 3-SO₂NH₂ I-6 4-CH₂NHSO₂NHCH₂CH₃ — H chloro — 4-SO₂NH₂ I-7  4-CH₂NHSO₂NHCH₂CH₃ — Hmethyl — 3-SO₂NH₂ I-8  4-CH₂NHSO₂NHCH₂CH₃ — H methyl — 4-SO₂NH₂ I-9 3-CH₂NHSO₂NHCH₂CH₃ — H chloro — 3-SO₂NH₂ I-10 3-CH₂NHSO₂NHCH₂CH₃ — Hchloro — 4-SO₂NH₂ I-11 3-CH₂NHSO₂NHCH₂CH₃ — H methyl — 3-SO₂NH₂ I-123-CH₂NHSO₂NHCH₂CH₃ — H methyl — 4-SO₂NH₂ I-13 3-CH₂NHSO₂NHCH₂CH₃ — Hfluoro — 3-SO₂NH₂ I-14 3-CH₂NHSO₂NHCH₂CH₃ — H fluoro — 4-SO₂NH₂ I-154-CH₂NHSO₂NHCH₂CH₃ 2-methyl H chloro — 3-SO₂NH₂ I-16 4-CH₂NHSO₂NHCH₂CH₃2-methyl H chloro — 4-SO₂NH₂ I-17 4-CH₂NHSO₂NHCH₂CH₃ 2-methyl H chloro3,5-dimethyl 4-SO₂NH₂ I-18

— H chloro — 3-SO₂NH₂ I-19

— H chloro — 4-SO₂NH₂ I-20

— H chloro 3,5-dimethyl 4-SO₂NH₂ I-21

— H methyl — 3-SO₂NH₂ I-22

— H methyl — 4-SO₂NH₂ I-23

— H methyl 3,5-dimethyl 4-SO₂NH₂ I-24

2-methyl H chloro — 3-SO₂NH₂ I-25

2-methyl H chloro — 4-SO₂NH₂ I-26

2-methyl H chloro 3,5-dimethyl 4-SO₂NH₂ I-27

— H methyl — 3-SO₂NHC(O)CH₂CH₃ I-28

— H methyl — 3-SO₂NHC(O)CH₂CH₃ I-29 3-CH₂NHSO₂NHCH₂CH₃ — H methyl4-methyl 3-SO₂NH₂ I-30 3-CH₂NHSO₂N(CH₃)₂ — H methyl — 3-SO₂NH₂ I-313-CH₂NHSO₂N(CH₃)₂ — H methyl — 4-SO₂NH₂ I-32 3-CH₂NHSO₂N(CH₃)₂ — Hmethyl 4-methyl 3-SO₂NH₂

TABLE II II

# R⁷ R⁸ Y R⁶ R² R¹ X R³ —SO₂N(R⁴)R⁵ II-1 Me Me —CH₂— —(CH₂)₂— — H F 4-Me3-SO₂NH₂ II-2 Me Me —CH₂— —CH₂— — H F 4-Me 3-SO₂NH₂ II-3 Me Me —CH₂——CH₂— — H Me — 4-SO₂NH₂ II-4 Me Me —CH₂— —CH₂— — H Me — 3-SO₂NH₂ II-5 MeMe —CH₂— —CH₂— — H Me 4-Me 3-SO₂NH₂

Prodrugs

Those of skill in the art will appreciate that the 2,4-pyrimidinediaminecompounds described herein can include functional groups that can bemasked with progroups to create prodrugs. Such prodrugs are usually, butneed not be, pharmacologically inactive until converted into theiractive drug form. Indeed, many of the 2,4-pyrimidinediamine compoundsdescribed in this invention include promoieties that are hydrolyzable orotherwise cleavable under conditions of use. For example, ester groupscommonly undergo acid-catalyzed hydrolysis to yield the parentcarboxylic acid when exposed to the acidic conditions of the stomach orbase-catalyzed hydrolysis when exposed to the basic conditions of theintestine or blood. Thus, when administered to a subject orally,2,4-pyrimidinediamine compounds that include ester moieties can beconsidered prodrugs of their corresponding carboxylic acid, regardlessof whether the ester form is pharmacologically active.

The mechanism by which the progroup(s) metabolizes is not critical andcan be caused, for example, by hydrolysis under the acidic conditions ofthe stomach, as described above, and/or by enzymes present in thedigestive tract and/or tissues or organs of the body. Indeed, theprogroup(s) can be selected to metabolize at a particular site withinthe body. For example, many esters are cleaved under the acidicconditions found in the stomach. Prodrugs designed to cleave chemicallyin the stomach to the active 2,4-pyrimidinediamine can employ progroupsincluding such esters. Alternatively, the progroups can be designed tometabolize in the presence of enzymes such as esterases, amidases,lipolases, and phosphatases, including ATPases and kinase, etc.Progroups including linkages capable of metabolizing in vivo are wellknown and include, by way of example and not limitation, ethers,thioethers, silylethers, silylthioethers, esters, thioesters,carbonates, thiocarbonates, carbamates, thiocarbamates, ureas,thioureas, and carboxamides. In some instances, a “precursor” group thatis oxidized by oxidative enzymes such as, for example, cytochrome P₄₅₀of the liver, to a metabolizable group, can be selected.

In the prodrugs, any available functional moiety can be masked with aprogroup to yield a prodrug. Functional groups within the2,4-pyrimidinediamine compounds that can be masked with progroups forinclusion in a promoiety include, but are not limited to, amines(primary and secondary), hydroxyls, sulfanyls (thiols), and carboxyls. Awide variety of progroups, as well as the resultant promoieties,suitable for masking functional groups in active 2,4-pyrimidinediaminecompounds to yield prodrugs are well-known in the art. For example, ahydroxyl functional group can be masked as a sulfonate, ester, orcarbonate promoiety, which can be hydrolyzed in vivo to provide thehydroxyl group. An amino functional group can be masked as an amide,carbamate, imine, urea, phosphenyl, phosphoryl, or sulfenyl promoiety,which can be hydrolyzed in vivo to provide the amino group. A carboxylgroup can be masked as an ester (including silyl esters and thioesters),amide, or hydrazide promoiety, which can be hydrolyzed in vivo toprovide the carboxyl group. Other specific examples of suitableprogroups and their respective promoieties will be apparent to those ofskill in the art. All of these progroups, alone or in combinations, canbe included in the prodrugs.

In some embodiments of the 2,4-pyrimidinediamine compounds and methodsof using the compounds, the progroup(s) can be attached to any availableprimary or secondary amine, including, for example, the N2 nitrogen atomof the 2,4-pyrimidinediamine, the N4 nitrogen atom of the2,4-pyrimidinediamine, and/or a primary or secondary nitrogen atomincluded in a substituent on the 2,4-pyrimidinediamine.

As noted above, the identity of the progroup is not critical, providedthat it can be metabolized under the desired conditions of use, forexample, under the acidic conditions found in the stomach and/or byenzymes found in vivo, to yield a biologically active group, e.g., the2,4-pyrimidinediamines as described herein. Thus, skilled artisans willappreciate that the progroup can include virtually any known orlater-discovered hydroxyl, amine or thiol protecting group. Non-limitingexamples of suitable protecting groups can be found, for example, inProtective Groups in Organic Synthesis, Greene & Wuts, 2^(nd) Ed., JohnWiley & Sons, New York, 1991 (especially pages 10-142 (alcohols, 277-308(thiols) and 309-405 (amines) the disclosure of which is incorporatedherein by reference).

Additionally, the identity of the progroup(s) can also be selected so asto impart the prodrug with desirable characteristics. For example,lipophilic groups can be used to decrease water solubility andhydrophilic groups can be used to increase water solubility. In thisway, prodrugs specifically tailored for selected modes of administrationcan be obtained. The progroup can also be designed to impart the prodrugwith other properties, such as, for example, improved passive intestinalabsorption, improved transport-mediated intestinal absorption,protection against fast metabolism (slow-release prodrugs),tissue-selective delivery, passive enrichment in target tissues, andtargeting-specific transporters. Groups capable of imparting prodrugswith these characteristics are well-known and are described, forexample, in Ettmayer et al., 2004, J. Med. Chem. 47(10):2393-2404, thedisclosure of which is incorporated by reference. All of the variousgroups described in these references can be utilized in the prodrugsdescribed herein.

A particularly useful progroup of the invention is —CH₂OP(OH)₂ as wellas esters, mixed acid esters and salts thereof. In some embodiments, the—CH₂OP(OH)₂ progroup is attached via an NH, annular or not, of theparent molecule. There can be more than one such progroup. Thus, oneembodiment is a compound of formula V,

or solvate thereof, where R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, X, p and q are asdefined in the summary and detailed description, R⁴⁰ is H or R⁵⁰, and atleast one of R⁴, R⁵, R⁶, R⁷, R⁸ and R⁴⁰ is R⁵⁰; where R⁵⁰ is—CH₂OP(O)(OR¹¹)₂; each is independently H, C₁₋₆alkyl or a monovalentcationic group, or two together with the atoms to which they areattached, form a 4-8 membered cyclic phosphate group

where each R⁵⁵ is independently H, optionally substituted C₁₋₆alkyl,optionally substituted 3-8 membered heteroalicyclyl, optionallysubstituted C₆₋₁₄aryl, optionally substituted C₇₋₂₀arylalkyl, optionallysubstituted 5-14 membered heteroaryl or optionally substituted 6-15membered heteroarylalkyl; z is 0, 1, 2 or 3; or two R¹¹ togetherrepresent a divalent cationic group selected from Ba²⁺, Bi²⁺, Ca²⁺,Cu²⁺, Mg²⁺, Ni²⁺, Sr²⁺ and Zn²⁺.

In one embodiment, the cyclic phosphate group is a 5 or 6-memberedcyclic phosphate group, where —CH₂OP(O)(OR¹¹)₂ is

or two R¹¹ together represent a divalent cationic group selected fromCa²⁺, Mg²⁺ and Zn²⁺.

Another embodiment is a compound of formula VI,

or solvate thereof, where R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, X, p and q are asdefined in the summary and detailed description, and R¹¹ is defined asabove.

While not intending to be bound by any particular theory of operation,it is believed that progroups —CH₂OP(O)(OR¹¹)₂, e.g according to formulaVI, metabolize to active compounds of the invention via thecorresponding hydroxymethylamine intermediate illustrated below:

Such hydroxymethylamine compounds, although isolable, are known to beunstable under physiological conditions and various pH ranges where theyhydrolyze in vivo to yield formaldehyde and the active drug substance.Based on this observation, compounds of the invention includehydroxymethyl progroups that can be metabolized in vivo, for example bythe acidic conditions of the stomach and/or by enzymes present in thedigestive tract or other organs and/or tissues or fluids with the body,to yield the active drug substance 2,4-pyrimidinediamine.

Moreover, it is expected that the amino and thio analogs of thesehydroxymethylamines, will be similarly unstable at physiologicalconditions and also hydrolyze in vivo to the active 2,4-pyrimdiendiaminedrug. Accordingly, compounds of the invention include thesecorresponding primary amino and thiol compounds. Also, the inventionincludes compounds in which the primary amine, thiol and hydroxy groupsare masked with “protecting” groups that are removed under physiologicalconditions of use to yield the corresponding hydroxymethyl, thiol methyland aminomethyl compounds, that is, with these “protecting groups” thesecompounds will likewise make suitable prodrugs.

Suitability of any particular progroup for a desired mode ofadministration can be confirmed in biochemical assays. For example, if aprodrug is to be administered by injection into a particular tissue ororgan and the identities of the various enzyme(s) expressed in thetissue or organ are known, the particular prodrug can be tested formetabolism in biochemical assays with the isolated enzyme(s).Alternatively, the particular prodrug can be tested for metabolism tothe active 2,4-pyrimidinediamine compound with tissue and/or organextracts. Using tissue and/or organ extracts can be of particularconvenience when the identity(ies) of the enzymes expressed in thetarget tissues or organs are unknown or in instances when the isolatedenzymes are not conveniently available. Skilled artisans will be able toreadily select progroups having metabolic properties (such as kinetics)suitable for particular applications using such in vitro tests. Specificprodrugs could also be tested for suitable metabolism in vitro animalmodels.

Compounds of the invention bearing the —CH₂OP(O)(OR¹¹)₂ progroup can besynthesized, e.g., as depicted below for compounds of formula VI, forexample, when appropriate protecting groups are installed elsewhere inthe molecule.

Typically, the proton on the 4-NH of the pyrimidinediamine system can beselectively alkylated over the 2-NH with the appropriate phosphonatereagent and reaction conditions, where LG is a suitable leaving group toform compounds of the invention, in this case of formula VI. Furtherdescription of how to make progroups of formula —CH₂OP(O)(OR¹¹)₂ asdescribed herein is specifically taught in U.S. Pat. No. 7,449,458,entitled “Pyrimidinediamine Prodrugs and their Uses,” the disclosure ofwhich is hereby incorporated by reference in its entirety.

Pharmaceutical Compositions

Another embodiment is a pharmaceutical composition including a compoundas described in any of the embodiments above. Pharmaceuticalcompositions described herein can be manufactured by means ofconventional mixing, dissolving, granulating, dragee-making levigating,emulsifying, encapsulating, entrapping, or lyophilization processes. Thecompositions can be formulated in conventional manner using one or morephysiologically acceptable carriers, diluents, excipients, orauxiliaries which facilitate processing of the active compounds intopreparations which can be used pharmaceutically.

The 2,4-pyrimidinediamine compound can be formulated in thepharmaceutical compositions per se, or in the form of a hydrate,solvate, N-oxide, or pharmaceutically acceptable salt, as describedherein. Typically, such salts are more soluble in aqueous solutions thanthe corresponding free acids and bases, but salts having lowersolubility than the corresponding free acids and bases can also beformed.

One embodiment is a pharmaceutical formulation including a compound offormula I, as described herein, or a prodrug thereof, and at least onepharmaceutically acceptable excipient, diluent, preservative,stabilizer, or mixture thereof.

The compounds can be provided in a variety of formulations and dosages.The compounds can be provided in a pharmaceutically acceptable form,including where the compound can be formulated in the pharmaceuticalcompositions per se, or in the form of a hydrate, solvate, N-oxide, orpharmaceutically acceptable salt, as described herein. Typically, suchsalts are more soluble in aqueous solutions than the corresponding freeacids and bases, but salts having lower solubility than thecorresponding free acids and bases can also be formed. It is to beunderstood that reference to the compound, 2,4-pyrimidinediaminecompound, or “active” in discussions of formulations is also intended toinclude, where appropriate as known to those of skill in the art,formulation of the prodrugs of the 2,4-pyrimidinediamine compounds. Inone embodiment, the compounds are provided as non-toxic pharmaceuticallyacceptable salts, as noted previously. Suitable pharmaceuticallyacceptable salts of the compounds described herein include acid additionsalts such as those formed with hydrochloric acid, fumaric acid,p-toluenesulphonic acid, maleic acid, succinic acid, acetic acid, citricacid, tartaric acid, carbonic acid, or phosphoric acid. Salts of aminegroups can also include quaternary ammonium salts in which the aminonitrogen atom carries a suitable organic group such as an alkyl,alkenyl, alkynyl, or substituted alkyl moiety. Furthermore, where thecompounds of the invention carry an acidic moiety, suitablepharmaceutically acceptable salts thereof can include metal salts suchas alkali metal salts, e.g., sodium or potassium salts; and alkalineearth metal salts, e.g., calcium or magnesium salts.

The pharmaceutical compositions for the administration of the2,4-pyrimidinediamine compounds can be conveniently presented in dosageunit form and can be prepared by any of the methods well known in theart of pharmacy. The pharmaceutical compositions can be, for example,prepared by uniformly and intimately bringing the active ingredient intoassociation with a liquid carrier, a finely divided solid carrier orboth, and then, if necessary, shaping the product into the desiredformulation. In the pharmaceutical composition the active objectcompound is included in an amount sufficient to produce the desiredtherapeutic effect.

The 2,4-pyrimidinediamine compounds can be administered by oral,parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV,intracisternal injection or infusion, subcutaneous injection, orimplant), by inhalation spray nasal, vaginal, rectal, sublingual,urethral (e.g., urethral suppository) or topical routes ofadministration (e.g., gel, ointment, cream, aerosol, etc.) and can beformulated, alone or together, in suitable dosage unit formulationscontaining conventional non-toxic pharmaceutically acceptable carriers,adjuvants, excipients, and vehicles appropriate for each route ofadministration. In addition to the treatment of warm-blooded animalssuch as mice, rats, horses, cattle, sheep, dogs, cats, and monkeys, thecompounds described herein can be effective in humans.

Administration of the compounds described herein, or theirpharmaceutically acceptable salts, in pure form or in an appropriatepharmaceutical composition, can be carried out via any of the acceptedmodes of administration or agents for serving similar utilities. Thus,administration can be, for example, orally, nasally, parenterally(intravenous, intramuscular, or subcutaneous), topically, transdermally,intravaginally, intravesically, intracistemally, or rectally, in theform of solid, semi-solid, lyophilized powder, or liquid dosage forms,such as for example, tablets, suppositories, pills, soft elastic andhard gelatin capsules, powders, solutions, suspensions, or aerosols, orthe like, preferably in unit dosage forms suitable for simpleadministration of precise dosages.

D. Methods of the Invention

The present invention provides 2,4-pyrimidinediamine compounds andprodrugs thereof, as described herein, for use in therapy for theconditions described herein. The present invention further provides useof the compounds of the present invention in the manufacture of amedicament for the treatment of conditions in which targeting of the JAKpathway or inhibition of JAK kinases, particularly JAK3, aretherapeutically useful. The methods include conditions where thefunction of lymphocytes, macrophages, or mast cells is involved.Conditions in which targeting of the JAK pathway or inhibition of theJAK kinases, particularly JAK3, are therapeutically useful includeleukemia, lymphoma, transplant rejection (e.g., pancreas islettransplant rejection), bone marrow transplant applications (e.g.,graft-versus-host disease)), autoimmune diseases (e.g., rheumatoidarthritis, etc.), inflammation (e.g., asthma, etc.) and other conditionsas described in greater detail herein.

As noted previously, numerous conditions can be treated using the2,4-substituted pyrimidinediamine compounds, prodrugs thereof, andmethods of treatment as described herein. As used herein, “Treating” or“treatment” of a disease in a patient refers to (1) preventing thedisease from occurring in a patient that is predisposed or does not yetdisplay symptoms of the disease; (2) inhibiting the disease or arrestingits development; or (3) ameliorating or causing regression of thedisease. As well understood in the art, “treatment” is an approach forobtaining beneficial or desired results, including clinical results. Forthe purposes of this invention, beneficial or desired results caninclude one or more, but are not limited to, alleviation or ameliorationof one or more symptoms, diminishment of extent of a condition,including a disease, stabilized (i.e., not worsening) state of acondition, including diseases, preventing spread of disease, delay orslowing of condition, including disease, progression, amelioration orpalliation of the condition, including disease, state, and remission(whether partial or total), whether detectable or undetectable.Preferred are compounds that are potent and can be administered locallyat very low doses, thus minimizing systemic adverse effects.

The compounds described herein are potent and selective inhibitors ofJAK kinases and are particularly selective for cytokine signalingpathways containing JAK3. As a consequence of this activity, thecompounds can be used in a variety of in vitro, in vivo, and ex vivocontexts to regulate or inhibit JAK kinase activity, signaling cascadesin which JAK kinases play a role, and the biological responses effectedby such signaling cascades. For example, in one embodiment, thecompounds can be used to inhibit JAK kinase, either in vitro or in vivo,in virtually any cell type expressing the JAK kinase, such as inhematopoietic cells in which, for example, JAK3 is predominantlyexpressed. They may also be used to regulate signal transductioncascades in which JAK kinases, particularly JAK3, play a role. SuchJAK-dependent signal transduction cascades include, but are not limitedto, the signaling cascades of cytokine receptors that involve the commongamma chain, such as, for example, the IL-4, IL-7, IL-5, IL-9, IL-15 andIL-21, or IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21 receptor signalingcascades. The compounds may also be used in vitro or in vivo toregulate, and in particular to inhibit, cellular or biological responsesaffected by such JAK-dependent signal transduction cascades. Suchcellular or biological responses include, but are not limited to,IL-4/ramos CD23 upregulation and IL-2 mediated T-cell proliferation.Importantly, the compounds can be used to inhibit JAK kinases in vivo asa therapeutic approach towards the treatment or prevention of diseasesmediated, either wholly or in part, by a JAK kinase activity (referredto herein as “JAK kinase mediated diseases”). Non-limiting examples ofJAK kinase mediated diseases that can be treated or prevented with thecompounds include, but are not limited to, the following: allergies;asthma; autoimmune diseases such as transplant rejection (e.g., kidney,heart, lung, liver, pancreas, skin; host versus graft reaction (HVGR),and graft versus host reaction (GVHR)), rheumatoid arthritis, andamyotrophic lateral sclerosis; T-cell mediated autoimmune diseases suchas multiple sclerosis, psoraiasis, and Sjogren's syndrome; Type IIinflammatory diseases such as vascular inflammation (includingvasculitis, arteritis, atherosclerosis, and coronary artery disease);diseases of the central nervous system such as stroke; pulmonarydiseases such as bronchitis obliteraus and primary pulmonaryhypertension; solid, delayed Type IV hypersensitivity reactions; andhematologic malignancies such as leukemia and lymphomas.

One embodiment is a method of inhibiting an activity of a JAK kinase,including contacting the JAK kinase with an amount of a compound,effective to inhibit an activity of the JAK kinase, of formula I:

wherein:

-   -   p is 0, 1, 2, or 3;    -   q is 0, 1, 2, or 3;    -   X is hydrogen, alkyl, substituted alkyl, hydroxy, alkoxy,        substituted alkoxy, amino, substituted amino, carboxyl, carboxyl        ester, —C(O)N(R⁴)R⁵, cyano, halo, nitro, alkenyl, substituted        alkenyl, alkynyl, substituted alkynyl, cycloalkynyl or        substituted cycloalkynyl;    -   Y is a straight or branched chain C₁₋₆ alkylene group,        cycloalkylene or substituted cycloalkylene;    -   R¹ is hydrogen, alkyl, substituted alkyl, cycloalkyl or        substituted cycloalkyl;    -   each R² independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, amino, substituted amino, aryl, substituted        aryl, aryloxy, substituted aryloxy, cyano, cycloalkyl,        substituted cycloalkyl, cycloalkoxy, substituted cycloalkoxy,        heteroaryl, substituted heteroaryl, heteroaryloxy, substituted        heteroaryloxy, heterocyclyl, substituted heterocyclyl,        heterocyclyloxy, substituted heterocyclyloxy, aminoacyl,        aminoacyloxy, carboxyl, carboxyl ester, carbonate ester,        —C(O)N(R⁴)R⁵, nitro or halo;    -   each R³ independently is alkyl, substituted alkyl, alkoxy,        substituted alkoxy, amino, substituted amino, aryl, substituted        aryl, aryloxy, substituted aryloxy, cyano, cycloalkyl,        substituted cycloalkyl, cycloalkoxy, substituted cycloalkoxy,        heteroaryl, substituted heteroaryl, heteroaryloxy, substituted        heteroaryloxy, heterocyclyl, substituted heterocyclyl,        heterocyclyloxy, substituted heterocyclyloxy, aminoacyl,        aminoacyloxy, carboxyl, carboxyl ester, carbonate ester, nitro        or halo;    -   each R⁴ independently is hydrogen, alkyl, substituted alkyl,        cycloalkyl, substituted cycloalkyl, heterocyclyl, substituted        heterocyclyl, carboxyl, carboxyl ester, aryl, substituted aryl,        heteroaryl, substituted heteroaryl, acyl or M⁺ wherein M⁺ is K⁺,        Na⁺, Li⁺ or ⁺N(R⁹)₄, wherein each R⁹ independently is hydrogen        or alkyl, and the nitrogen immediately adjacent to R⁵ is        anionic; or        -   R⁴ and R⁵ together with the intervening atom or atoms bound            thereto, form a heterocyclyl or substituted heterocyclyl            group;    -   each R⁵ independently is hydrogen, alkyl, substituted alkyl,        amino, cycloalkyl, substituted cycloalkyl, heterocyclyl,        substituted heterocyclyl, aryl, substituted aryl, heteroaryl,        substituted heteroaryl, carboxyl, carboxyl ester or acyl;    -   R⁶ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl; or    -   R⁶ is a straight or branched chain C₁₋₆alkylene group,        cycloalkylene or substituted cycloalkylene, linking the nitrogen        bearing R⁶ and the ring bearing Y    -   R⁷ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl;    -   R⁸ is hydrogen, alkyl, substituted alkyl, cycloalkyl,        substituted cycloalkyl, heterocyclyl, substituted heterocyclyl,        aryl, substituted aryl, heteroaryl, substituted heteroaryl or        acyl; or    -   R⁷ and R⁸ together, with nitrogen to which they are bound, form        a heterocyclyl or substituted heterocyclyl group;    -   wherein at least one of R⁷ and R⁸ is non-hydrogen; and        each of R⁶, R⁷, and R⁸ optionally are M⁺, wherein M⁺ is K⁺, Na⁺,        Li⁺ or ⁺N(R⁹)₄, wherein each R⁹ independently is hydrogen or        alkyl, and the nitrogen bearing M⁺ is anionic.

In the various method embodiments below, when the compound of Formula Iis referred to, also meant to be included are distinct and analogousembodiments that apply to compounds of formula II, III and W, as well asto species I-1 through I-32 and II-1 through II-5.

In another embodiment, this invention provides a method of inhibiting anactivity of a JAK kinase, including contacting the JAK kinase with anamount of a compound effective to inhibit an activity of the JAK kinase,where the compound is according to formula I, as described herein. Incertain embodiments of the methods described herein, the method iscarried out in vivo.

In another embodiment, this invention provides a method of inhibiting anactivity of a JAK kinase, including contacting in vitro a JAK3 kinasewith an amount of a compound effective to inhibit an activity of the JAKkinase, where the compound is according to formula I, as describedherein.

In a specific embodiment, the compounds can be used to treat and/orprevent rejection in organ and/or tissue transplant recipients (i.e.,treat and/or prevent allograft rejection). Allografts can be rejectedthrough either a cell-mediated or humoral immune reaction of therecipient against transplant (histocompability) antigens present on themembranes of the donor's cells. The strongest antigens are governed by acomplex of genetic loci termed human leukocyte group A (HLA) antigens.Together with the ABO blood groups antigens, they are the chieftransplantation antigens detectable in humans.

Rejection following transplantation can generally be broken into threecategories: hyperacute, occurring hours to days followingtransplantation; acute, occurring days to months followingtransplantation; and chronic, occurring months to years followingtransplantation.

Hyperacute rejection is caused mainly by the production of hostantibodies that attack the graft tissue. In a hyperacute rejectionreaction, antibodies are observed in the transplant vascular very soonafter transplantation. Shortly thereafter, vascular clotting occurs,leading to ischemia, eventual necrosis and death. The graft infarctionis unresponsive to known immunosuppressive therapies. Because HLAantigens can be identified in vitro, pre-transplant screening is used tosignificantly reduce hyperacute rejection. As a consequence of thisscreening, hyperacute rejection is relatively uncommon today.

Acute rejection is thought to be mediated by the accumulation of antigenspecific cells in the graft tissue. The T-cell-mediated immune reactionagainst these antigens (i.e., HVGR or GVHR) is the principle mechanismof acute rejection. Accumulation of these cells leads to damage of thegraft tissue. It is believed that both CD4+ helper T-cells and CD8+cytotoxic T-cells are involved in the process and that the antigen ispresented by donor and host dendritic cells. The CD4+ helper T-cellshelp recruit other effector cells, such as macrophapges and eosinophils,to the graft. Accessing T-cell activation signal transduction cascades(for example, CD28, CD40L, and CD2 cascades) are also involved.

The cell-mediated acute rejection can be reversed in many cases byintensifying immunotherapy. After successful reversal, severely damagedelements of the graft heal by fibrosis and the remainder of the graftappears normal. After resolution of acute rejection, dosages ofimmunosuppressive drugs can be reduced to very low levels.

Chronic rejection, which is a particular problem in renal transplants,often progresses insidiously despite increased immunosuppressivetherapy. It is thought to be due, in large part, to cell-mediated Type Whypersensitivity. The pathologic profile differs from that of acuterejection. The arterial endothelium is primarily involved with extensiveproliferation that may gradually occlude the vessel lumen, leading toischemia, fibrosis, a thickened intima, and atherosclerotic changes.Chronic rejection is mainly due to a progressive obliteration of graftvasculature and resembles a slow, vasculitic process.

In Type IV hypersensitivity, CD8 cytotoxic T-cells and CD4 helper Tcells recognize either intracellular or extracellular synthesizedantigen when it is complexed, respectively, with either Class I or ClassII MHC molecules. Macrophages function as antigen-presenting cells andrelease IL-1, which promotes proliferation of helper T-cells. HelperT-cells release interferon gamma and IL-2, which together regulatedelayed hyperactivity reactions mediated by macrophage activation andimmunity mediated by T cells. In the case of organ transplant, thecytotoxic T-cells destroy the graft cells on contact.

Since JAK kinases play a critical role in the activation of T-cells, the2,4-pyrimidinediamine compounds described herein can be used to treatand/or prevent many aspects of transplant rejection, and areparticularly useful in the treatment and/or prevention of rejectionreactions that are mediated, at least in part, by T-cells, such as HVGRor GVHR. The 2,4-pyrimidinediamine compounds can also be used to treatand/or prevent chronic rejection in transplant recipients and, inparticular, in renal transplant recipients. The compound can also beadministered to a tissue or an organ prior to transplanting the tissueor organ in the transplant recipient.

In another embodiment, this invention provides a method of treating aT-cell mediated autoimmune disease, including administering to a patientsuffering from such an autoimmune disease an amount of a compoundeffective to treat the autoimmune disease where the compound isaccording to formula I, as described herein. In certain embodiments ofthe methods the autoimmune disease is multiple sclerosis (MS),psoriasis, or Sjogran's syndrome.

Therapy using the 2,4-pyrimidinediamine compounds described herein canbe applied alone, or it can be applied in combination with or adjunctiveto other common immunosuppressive therapies, such as, for example, thefollowing: mercaptopurine; corticosteroids such as prednisone;methylprednisolone and prednisolone; alkylating agents such ascyclophosphamide; calcineurin inhibitors such as cyclosporine,sirolimus, and tacrolimus; inhibitors of inosine monophosphatedehydrogenase (IMPDH) such as mycophenolate, mycophenolate mofetil, andazathioprine; and agents designed to suppress cellular immunity whileleaving the recipient's humoral immunologic response intact, includingvarious antibodies (for example, antilymphocyte globulin (ALG),antithymocyte globulin (ATG), monoclonal anti-T-cell antibodies (OKT3))and irradiation. These various agents can be used in accordance withtheir standard or common dosages, as specified in the prescribinginformation accompanying commercially available forms of the drugs (seealso: the prescribing information in the 2006 Edition of The Physician'sDesk Reference), the disclosures of which are incorporated herein byreference. Azathioprine is currently available from SalixPharmaceuticals, Inc., under the brand name AZASAN; mercaptopurine iscurrently available from Gate Pharmaceuticals, Inc., under the brandname PURINETHOL; prednisone and prednisolone are currently availablefrom Roxane Laboratories, Inc.; Methyl prednisolone is currentlyavailable from Pfizer; sirolimus (rapamycin) is currently available fromWyeth-Ayerst under the brand name RAPAMUNE; tacrolimus is currentlyavailable from Fujisawa under the brand name PROGRAF; cyclosporine iscurrent available from Novartis under the brand name SANDIMMUNE and fromAbbott under the brand name GENGRAF; IMPDH inhibitors such asmycophenolate mofetil and mycophenolic acid are currently available fromRoche under the brand name CELLCEPT and from Novartis under the brandname MYFORTIC; azathioprine is currently available from Glaxo SmithKline under the brand name IMURAN; and antibodies are currentlyavailable from Ortho Biotech under the brand name ORTHOCLONE, fromNovartis under the brand name SIMULECT (basiliximab), and from Rocheunder the brand name ZENAPAX (daclizumab).

In another embodiment, the 2,4-pyrimidinediamine compounds could beadministered either in combination or adjunctively with an inhibitor ofa Syk kinase. Syk kinase is a tyrosine kinase known to play a criticalrole in Fcγ receptor signaling, as well as in other signaling cascades,such as those involving B-Cell receptor signaling (Turner et al.,(2000), Immunology Today 21:148-154) and integrins beta(1), beta (2),and beta (3) in neutrophils (Mocsavi et al., (2002), Immunity16:547-558). For example, Syk kinase plays a pivotal role in highaffinity IgE receptor signaling in mast cells that leads to activationand subsequent release of multiple chemical mediators that triggerallergic attacks. However, unlike the JAK kinases, which help regulatethe pathways involved in delayed or cell-mediated Type IVhypersensitivity reactions, Syk kinase helps regulate the pathwaysinvolved in immediate IgE-mediated, Type I hypersensitivity reactions.Certain compounds that affect the Syk pathway may or may not also affectthe JAK pathways.

Suitable Syk inhibitory compounds are described, for example, in Ser.No. 10/355,543 filed Jan. 31, 2003 (publication no. 2004/0029902); WO03/063794; Ser. No. 10/631,029 filed Jul. 29, 2003 (publication no.2007/0060603); WO 2004/014382; Ser. No. 10/903,263 filed Jul. 30, 2004(publication no. 2005/0234049); PCT/US2004/24716 filed Jul. 30, 2004(WO005/016893); Ser. No. 10/903,870 filed Jul. 30, 2004 (publication no.2005/0209224); PCT/US2004/24920 filed Jul. 30, 2004; Ser. No. 60/630,808filed Nov. 24, 2004; Ser. No. 60/645,424 filed Jan. 19, 2005; and Ser.No. 60/654,620, filed Feb. 18, 2005, the disclosures of which areincorporated herein by reference. The 2,4-pyrimidinediamine describedherein and Syk inhibitory compounds could be used alone or incombination with one or more conventional transplant rejectiontreatments, as described above.

In a specific embodiment, the 2,4-pyrimidinediamine compounds can beused to treat or prevent these diseases in patients that are eitherinitially non-responsive (resistant) to or that become non-responsive totreatment with a Syk inhibitory compound or one of the other currenttreatments for the particular disease. The 2,4-pyrimidinediaminecompounds could also be used in combination with Syk inhibitorycompounds in patients that are Syk-compound resistant or non-responsive.Suitable Syk-inhibitory compounds with which the 2,4-pyrimidinediaminecompounds can be administered are provided supra.

In another embodiment, this invention provides a method of treating aT-cell mediated autoimmune disease, including administering to a patientsuffering from such an autoimmune disease an amount of a compoundaccording to formula I, in combination with or adjunctively to acompound that inhibits Syk kinase with an IC₅₀ of at least 10 μM,effective to treat the autoimmune disease.

In another embodiment, this invention provides a method of treating orpreventing allograft transplant rejection, either acute or chronic, in atransplant recipient, including administering to the transplantrecipient an amount of a compound according to formula I effective totreat or prevent the rejection. In a further embodiment, the compound isadministered to a tissue or an organ prior to or concurrent with,transplanting the tissue or organ in the transplant recipient. Inanother embodiment, the compound is administered to the tissue or organand the patient. In a specific embodiment the allograft transplantrejection is mediated by HVGR or GVHR. In another embodiment, theallograft transplant organ is a kidney, a heart, a liver, or a lung. Inanother embodiment, in which the allograft transplant organ is a kidney,a heart, a liver, or a lung, the compound is administered in combinationwith or adjunctively to another immunosuppressant. In a more specificembodiment, the immunosuppressant is cyclosporine, tacrolimus,sirolimus, an inhibitor of IMPDH, mycophenolate, mycophanolate mofetil,an anti-T-Cell antibody or OKT3.

The 2,4-pyrimidinediamine compounds described herein are cytokinemoderators of IL-4 signaling. As a consequence, the2,4-pyrimidinediamine compounds could slow the response of Type Ihypersensitivity reactions. Thus, in a specific embodiment, the2,4-pyrimidinediamine compounds could be used to treat such reactionsand, therefore, the diseases associated with, mediated by, or caused bysuch hypersensitivity reactions (for example, allergies),prophylactically. For example, an allergy sufferer could take one ormore of the JAK selective compounds described herein prior to expectedexposure to allergens to delay the onset or progress of, or eliminatealtogether, an allergic response.

When used to treat or prevent such diseases, the 2,4-pyrimidinediaminecompounds can be administered singly, as mixtures of one or more2,4-pyrimidinediamine compounds, or in mixture or combination with otheragents useful for treating such diseases and/or the symptoms associatedwith such diseases. The 2,4-pyrimidinediamine compounds can also beadministered in mixture or in combination with agents useful to treatother disorders or maladies, such as steroids, membrane stabilizers,5-lipoxygenase (5LO) inhibitors, leukotriene synthesis and receptorinhibitors, inhibitors of IgE isotype switching or IgE synthesis, IgGisotype switching or IgG synthesis, β-agonists, tryptase inhibitors,aspirin, cyclooxygenase (COX) inhibitors, methotrexate, anti-TNF drugs,rituximab, PD4 inhibitors, p38 inhibitors, PDE4 inhibitors, andantihistamines, to name a few. The 2,4-pyrimidinediamine compounds canbe administered per se in the form of prodrugs or as pharmaceuticalcompositions, including an active compound.

In another embodiment, this invention provides a method of treating orpreventing a Type IV hypersensitivity reaction, including administeringto a subject an amount of a compound effective to treat or prevent thehypersensitivity reaction, where the compound is according to formula I,as described herein. In one embodiment, the method is practicedprophylactically. In some embodiments, the compound is administeredprior to exposure to an allergen.

In another embodiment, this invention provides a method of inhibiting asignal transduction cascade in which JAK3 kinase plays a role, includingcontacting a cell expressing a receptor involved in such a signalingcascade with a compound, where the compound is according to formula I,as described herein.

In another embodiment, this invention provides a method of treating orpreventing a JAK kinase-mediated disease, including administering to asubject an amount of compound effective to treat or prevent the JAKkinase-mediated disease, where the compound is according to formula I,as described herein.

In another embodiment, this invention provides a method of treating orpreventing a JAK kinase-mediated disease, in which the JAK-mediateddisease is HVGR or GVHR, including administering to a subject an amountof compound effective to treat or prevent the JAK kinase-mediateddisease, where the compound is according to formula I, as describedherein.

In another embodiment, ocular disorders are treated using an effectiveamount of a compound of formula I, as described herein. In one aspect ofthe disclosed method for treating ocular disorders, administration ofone or more of the presently disclosed 2,4-pyrimidinediamine compoundsis effective to increase tear production volume as compared to untreatedtear production volume, thereby ameliorating a symptom of dry eyesyndrome. In one aspect, tear production volume is increased within fivedays, such as in less than four days, and in some examples in less thantwo days. In one embodiment, tear production volume is increased by atleast about 25% over initial tear production within two days of initialtreatment with a presently disclosed 2,4-pyrimidinediamine compound. Inother embodiments, tear production is increased at least about 30%, suchas at least about 50% over initial tear production within less than twodays. Increases in tear production upon administration of the presentcompounds results, in some instances, in tear production volumecomparable to normal tear production. Typically the disclosed compounds,when used for treating ocular disorders topically, are administered atleast once daily and typically at most twice a day.

As mentioned, another embodiment provides a method of treating a diseaseand/or disorder of the eye, which includes administering to a subject anamount of a compound effective to treat the disease and/or disorder ofthe eye wherein the compound is according to formula I, as describedherein. Diseases and disorders of the eye include, but are not limitedto, dry eye syndrome, uveitis, allergic conjunctivitus, glaucoma androsacea (of the eye). Dry eye syndrome (DES), otherwise known askeratoconjunctivitis sicca (KCS), keratitis sicca, sicca syndrome, orxerophthalmia, is an eye disease caused by decreased tear production orincreased tear film evaporation commonly found in humans and someanimals. Uveitis or iridocyclitis refers to inflammation of the middlelayer of the eye (the “uvea”) and in common usage may refer to anyinflammatory process involving the interior of the eye. Allergicconjunctivitis is inflammation of the conjunctiva (the membrane coveringthe white part of the eye) due to allergy. Glaucoma refers to a group ofdiseases that affect the optic nerve and involves a loss of retinalganglion cells in a characteristic pattern, i.e., a type of opticneuropathy. Raised intraocular pressure is a significant risk factor fordeveloping glaucoma (above 22 mmHg or 2.9 kPa), and inflammatoryprocesses, e.g uveitis, can cause this rise in intraocular pressure.Rosacea is a chronic inflammatory condition characterized by facialerythema but it can affect the eyes. As mentioned, compounds describedherein may be used to treat inflammatory responses. While not wishing tobe bound by theory, it is believed that compounds described herein areeffective treatments of these eye disorders due, at least in part, totheir JAK inhibitory activity.

Active compounds described herein typically inhibit the JAK/Statpathway. The activity of a specified compound as an inhibitor of a JAKkinase can be assessed in vitro or in vivo. In some embodiments, theactivity of a specified compound can be tested in a cellular assay.

Suitable assays include assays that determine inhibition of either thephosphorylation activity or ATPase activity of a JAK kinase. Thus, acompound is said to inhibit an activity of a JAK kinase if it inhibitsthe phosphorylation or ATPase activity of a JAK kinase with an IC₅₀ ofabout 20 μM or less.

“Cell proliferative disorder” refers to a disorder characterized byabnormal proliferation of cells. A proliferative disorder does not implyany limitation with respect to the rate of cell growth, but merelyindicates loss of normal controls that affect growth and cell division.Thus, in some embodiments, cells of a proliferative disorder can havethe same cell division rates as normal cells but do not respond tosignals that limit such growth. Within the ambit of “cell proliferativedisorder” is neoplasm or tumor, which is an abnormal growth of tissue.Cancer refers to any of various malignant neoplasms characterized by theproliferation of cells that have the capability to invade surroundingtissue and/or metastasize to new colonization sites.

“Hematopoietic neoplasm” refers to a cell proliferative disorder arisingfrom cells of the hematopoietic lineage. Generally, hematopoiesis is thephysiological process whereby undifferentiated cells or stem cellsdevelop into various cells found in the peripheral blood. In the initialphase of development, hematopoietic stem cells, typically found in thebone marrow, undergo a series of cell divisions to form multipotentprogenitor cells that commit to two main developmental pathways: thelymphoid lineage and the myeloid lineage. The committed progenitor cellsof the myeloid lineage differentiate into three major sub-branchesincluding the erythroid, megakaryocyte, and granulocyte/monocytedevelopmental pathways. An additional pathway leads to formation ofdendritic cells, which are involved in antigen presentation. Theerythroid lineage gives rise to red blood cells while the megakaryocyticlineage gives rise to blood platelets. Committed cells of thegranulocyte/monocyte lineage split into granulocyte or monocytedevelopmental pathways, the former pathway leading to formation ofneutrophils, eosinophils, and basophils and the latter pathway givingrise to blood monocytes and macrophages.

Committed progenitor cells of the lymphoid lineage develop into the Bcell pathway, T cell pathway, or the non-T/B cell pathway. Similar tothe myeloid lineage, an additional lymphoid pathway appears to give riseto dendritic cells involved in antigen presentation. The B cellprogenitor cell develops into a precursor B cell (pre-B), whichdifferentiates into B cells responsible for producing immunoglobulins.Progenitor cells of the T cell lineage differentiate into precursor Tcells (pre-T) that, based on the influence of certain cytokines, developinto cytotoxic or helper/suppressor T cells involved in cell mediatedimmunity. Non-T/B cell pathway leads to generation of natural killer(NK) cells. Neoplasms of hematopoietic cells can involve cells of anyphase of hematopoiesis, including hematopoietic stem cells, multipotentprogenitor cells, oligopotent committed progenitor cells, precursorcells, and mature differentiated cells. The categories of hematopoieticneoplasms can generally follow the descriptions and diagnostic criteriaemployed by those of skill in the art (see, e.g., InternationalClassification of Disease and Related Health Problems (ICD 10), WorldHealth Organization (2003)). Hematopoietic neoplasms can also becharacterized based on the molecular features, such as cell surfacemarkers and gene expression profiles, cell phenotype exhibited by theaberrant cells, and/or chromosomal aberrations (e.g., deletions,translocations, insertions, etc.) characteristic of certainhematopoietic neoplasms, such as the Philadelphia chromosome found inchronic myelogenous leukemia. Other classifications include NationalCancer Institute Working Formulation (Cancer, 1982, 49:2112-2135) andRevised European-American Lymphoma Classification (REAL).

“Lymphoid neoplasm” refers a proliferative disorder involving cells ofthe lymphoid lineage of hematopoiesis. Lymphoid neoplasms can arise fromhematopoietic stem cells as well as lymphoid committed progenitor cells,precursor cells, and terminally differentiated cells. These neoplasmscan be subdivided based on the phenotypic attributes of the aberrantcells or the differentiated state from which the abnormal cells arise.Subdivisions include, among others, B cell neoplasms, T cell neoplasms,NK cell neoplasms, and Hodgkin's lymphoma.

“Myeloid neoplasm” refers to proliferative disorder of cells of themyeloid lineage of hematopoiesis. Neoplasms can arise from hematopoieticstem cells, myeloid committed progenitor cells, precursor cells, andterminally differentiated cells. Myeloid neoplasms can be subdividedbased on the phenotypic attributes of the aberrant cells or thedifferentiated state from which the abnormal cells arise. Subdivisionsinclude, among others, myeloproliferative diseases,myelodysplastic/myeloproliferative diseases, myelodysplastic syndromes,acute myeloid leukemia, and acute biphenotypic leukemia.

Generally cell proliferative disorders treatable with the compoundsdisclosed herein relate to any disorder characterized by aberrant cellproliferation. These include various tumors and cancers, benign ormalignant, metastatic or non-metastatic. Specific properties of cancers,such as tissue invasiveness or metastasis, can be targeted using themethods described herein. Cell proliferative disorders include a varietyof cancers, including, among others, breast cancer, ovarian cancer,renal cancer, gastrointestinal cancer, kidney cancer, bladder cancer,pancreatic cancer, lung squamous carcinoma, and adenocarcinoma. Morespecifically, related to particular tissues, organs or areas of thebody, Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma,liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung:bronchogenic carcinoma (squamous cell, undifferentiated small cell,undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar)carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatoushanlartoma, inesothelioma; Gastrointestinal: esophagus (squamous cellcarcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach(carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma,insulinorna, glucagonoma, gastrinoma, carcinoid tumors, vipoma), smallbowel (adenocarcinorna, lymphoma, carcinoid tumors, Karposi's sarcoma,leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel(adenocarcinoma, tubular adenoma, villous adenoma, hamartoma,leiomyoma); Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor[nephroblastoma], lymphoma, leukemia), bladder and urethra (squamouscell carcinoma, transitional cell carcinoma, adenocarcinoma), prostateadenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonalcarcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cellcarcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver:hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma,angiosarcoma, hepatocellular adenoma, hemangioma; Bone: osteogenicsarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma,chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cellsarcoma), multiple myeloma, malignant giant cell tumor chordoma,osteochronfroma (osteocartilaginous exostoses), benign chondroma,chondroblastoma, chondromyxofibroma, osteoid osteoma and giant celltumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma,osteitis defomians), meninges (meningioma, meningiosarcoma,gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma,germinoma [pinealoma], glioblastoma multiform, oligodendroglioma,schwannoma, retinoblastoma, congenital tumors), spinal cordneurofibroma, meningioma, glioma, sarcoma); Gynecological: uterus(endometrial carcinoma), cervix (cervical carcinoma, pre-tumor cervicaldysplasia), ovaries (ovarian carcinoma [serous cystadenocarcinoma,mucinous cystadenocarcinoma, unclassified carcinoma], granulosa-thecalcell tumors, SertoliLeydig cell tumors, dysgerminoma, malignantteratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma,adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma,squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma],fallopian tubes (carcinoma); Hematologic: blood (myeloid leukemia [acuteand chronic], acute lymphoblastic leukemia, chronic lymphocyticleukemia, myeloproliferative diseases, multiple myeloma, myelodysplasticsyndrome), Hodgkin's disease, non-Hodgkin's lymphoma [malignantlymphoma]; Skin: malignant melanoma, basal cell carcinoma, squamous cellcarcinoma, Karposi's sarcoma, moles dysplastic nevi, lipoma, angioma,dermatofibroma, keloids, psoriasis; and Adrenal lands: neuroblastoma.The term “cancerous cell” as provided herein, includes a cell afflictedby any one of the above-identified conditions.

In some embodiments, the cell proliferative disorder treated is ahematopoietic neoplasm, which is aberrant growth of cells of thehematopoietic system. Hematopoietic malignancies can have its origins inpluripotent stem cells, multipotent progenitor cells, oligopotentcommitted progenitor cells, precursor cells, and terminallydifferentiated cells involved in hematopoiesis. Some hematologicalmalignancies are believed to arise from hematopoietic stem cells, whichhave the ability for self renewal. For instance, cells capable ofdeveloping specific subtypes of acute myeloid leukemia (AML) upontransplantation display the cell surface markers of hematopoietic stemcells, implicating hematopoietic stem cells as the source of leukemiccells. Blast cells that do not have a cell marker characteristic ofhematopoietic stem cells appear to be incapable of establishing tumorsupon transplantation (Blake et al., 1997, Blood 89:3104-3112). The stemcell origin of certain hematological malignancies also finds support inthe observation that specific chromosomal abnormalities associated withparticular types of leukemia can be found in normal cells ofhematopoietic lineage as well as leukemic blast cells. For instance, thereciprocal translocation t(9q34; 22q11) associated with approximately95% of chronic myelogenous leukemia appears to be present in cells ofthe myeloid, erythroid, and lymphoid lineage, suggesting that thechromosomal aberration originates in hematopoietic stem cells. Asubgroup of cells in certain types of CML displays the cell markerphenotype of hematopoietic stem cells.

Although hematopoietic neoplasms often originate from stem cells,committed progenitor cells or more terminally differentiated cells of adevelopmental lineage can also be the source of some leukemias. Forexample, forced expression of the fusion protein Bcr/Abl (associatedwith chronic myelogenous leukemia) in common myeloid progenitor orgranulocyte/macrophage progenitor cells produces a leukemic-likecondition. Moreover, some chromosomal aberrations associated withsubtypes of leukemia are not found in the cell population with a markerphenotype of hematopoietic stem cells, but are found in a cellpopulation displaying markers of a more differentiated state of thehematopoietic pathway (Turhan et al., 1995, Blood 85:2154-2161). Thus,while committed progenitor cells and other differentiated cells may haveonly a limited potential for cell division, leukemic cells may haveacquired the ability to grow unregulated, in some instances mimickingthe self-renewal characteristics of hematopoietic stem cells (Passegueet al., Proc. Natl. Acad. Sci. USA, 2003, 100:11842-9).

In some embodiments, the hematopoietic neoplasm treated is a lymphoidneoplasm, where the abnormal cells are derived from and/or display thecharacteristic phenotype of cells of the lymphoid lineage. Lymphoidneoplasms can be subdivided into B-cell neoplasms, T and NK-cellneoplasms, and Hodgkin's lymphoma. B-cell neoplasms can be furthersubdivided into precursor B-cell neoplasm and mature/peripheral B-cellneoplasm. Exemplary B-cell neoplasms are precursor B-lymphoblasticleukemia/lymphoma (precursor B-cell acute lymphoblastic leukemia) whileexemplary mature/peripheral B-cell neoplasms are B-cell chroniclymphocytic leukemia/small lymphocytic lymphoma, B-cell prolymphocyticleukemia, lymphoplasmacytic lymphoma, splenic marginal zone B-celllymphoma, hairy cell leukemia, plasma cell myeloma/plasmacytoma,extranodal marginal zone B-cell lymphoma of MALT type, nodal marginalzone B-cell lymphoma, follicular lymphoma, mantle-cell lymphoma, diffuselarge B-cell lymphoma, mediastinal large B-cell lymphoma, primaryeffusion lymphoma, and Burkitt's lymphoma/Burkitt cell leukemia. T-celland Nk-cell neoplasms are further subdivided into precursor T-cellneoplasm and mature (peripheral) T-cell neoplasms. Exemplary precursorT-cell neoplasm is precursor T-lymphoblastic lymphoma/leukemia(precursor T-cell acute lymphoblastic leukemia) while exemplary mature(peripheral) T-cell neoplasms are T-cell prolymphocytic leukemia T-cellgranular lymphocytic leukemia, aggressive NK-cell leukemia, adult T-celllymphoma/leukemia (HTLV-1), extranodal NK/T-cell lymphoma, nasal type,enteropathy-type T-cell lymphoma, hepatosplenic gamma-delta T-celllymphoma, subcutaneous panniculitis-like T-cell lymphoma, Mycosisfungoides/Sezary syndrome, Anaplastic large-cell lymphoma, T/null cell,primary cutaneous type, Peripheral T-cell lymphoma, not otherwisecharacterized, Angioimmunoblastic T-cell lymphoma, Anaplastic large-celllymphoma, T/null cell, primary systemic type. The third member oflymphoid neoplasms is Hodgkin's lymphoma, also referred to as Hodgkin'sdisease. Exemplary diagnosis of this class that can be treated with thecompounds include, among others, nodular lymphocyte-predominantHodgkin's lymphoma, and various classical forms of Hodgkin's disease,exemplary members of which are Nodular sclerosis Hodgkin's lymphoma(grades 1 and 2), Lymphocyte-rich classical Hodgkin's lymphoma, Mixedcellularity Hodgkin's lymphoma, and Lymphocyte depletion Hodgkin'slymphoma. In various embodiments, any of the lymphoid neoplasms that areassociated with aberrant JAK activity can be treated with the JAKinhibitory compounds.

In some embodiments, the hematopoietic neoplasm treated is a myeloidneoplasm. This group includes a large class of cell proliferativedisorders involving or displaying the characteristic phenotype of thecells of the myeloid lineage. Myeloid neoplasms can be subdivided intomyeloproliferative diseases, myelodysplastic/myeloproliferativediseases, myelodysplastic syndromes, and acute myeloid leukemias.Exemplary myeloproliferative diseases are chronic myelogenous leukemia(e.g., Philadelphia chromosome positive (t(9; 22)(qq34; q11)), chronicneutrophilic leukemia, chronic eosinophilic leukemia/hypereosinophilicsyndrome, chronic idiopathic myelofibrosis, polycythemia vera, andessential thrombocythemia. Exemplary myelodysplastic/myeloproliferativediseases are chronic myelomonocytic leukemia, atypical chronicmyelogenous leukemia, and juvenile myelomonocytic leukemia. Exemplarymyelodysplastic syndromes are refractory anemia, with ringedsideroblasts and without ringed sideroblasts, refractory cytopenia(myelodysplastic syndrome) with multilineage dysplasia, refractoryanemia (myelodysplastic syndrome) with excess blasts, 5q-syndrome, andmyelodysplastic syndrome. In various embodiments, any of the myeloidneoplasms that are associated with aberrant JAK activity can be treatedwith the JAK inhibitory compounds.

In some embodiments, the JAK inhibitory compounds can be used to treatAcute myeloid leukemias (AML), which represent a large class of myeloidneoplasms having its own subdivision of disorders. These subdivisionsinclude, among others, AMLs with recurrent cytogenetic translocations,AML with multilineage dysplasia, and other AML not otherwisecategorized. Exemplary AMLs with recurrent cytogenetic translocationsinclude, among others, AML with t(8; 21)(q22; q22), AML1(CBF-alpha)/ETO,Acute promyelocytic leukemia (AML with t(15; 17)(q22; q11-12) andvariants, PML/RAR-alpha), AML with abnormal bone marrow eosinophils(inv(16)(p13q22) or t(16; 16)(p13; q11), CBFb/MYH11X), and AML with11q23 (MLL) abnormalities. Exemplary AML with multilineage dysplasia arethose that are associated with or without prior myelodysplasticsyndrome. Other acute myeloid leukemias not classified within anydefinable group include, AML minimally differentiated, AML withoutmaturation, AML with maturation, Acute myelomonocytic leukemia, Acutemonocytic leukemia, Acute erythroid leukemia, Acute megakaryocyticleukemia, Acute basophilic leukemia, and Acute panmyelosis withmyelofibrosis.

One means of assaying for such inhibition is detection of the effect ofthe 2,4-pyrimidinediamine compounds on the upregulation of downstreamgene products. In the Ramos/IL4 assay, B-cells are stimulated with thecytokine Interleukin-4 (IL-4) leading to the activation of the JAK/Statpathway through phosphorylation of the JAK family kinases, JAK1 andJAK3, which in turn phosphorylate and activate the transcription factorStat-6. One of the genes upregulated by activated Stat-6 is the lowaffinity IgE receptor, CD23. To study the effect of inhibitors (e.g.,the 2,4-substituted pyrimindinediamine compounds described herein) onthe JAK1 and JAK3 kinases, human Ramos B cells are stimulated with humanIL-4. 20 to 24 hours post stimulation, cells are stained forupregulation of CD23 and analyzed using flow cytometry (FACS). Areduction of the amount of CD23 present compared to control conditionsindicates the test compound actively inhibits the JAK kinase pathway.

The activity of the compounds described herein can further becharacterized by assaying the effect of the 2,4-pyrimidinediaminecompounds described herein on the proliferative response of primaryhuman T-cells. In this assay, primary human T-cells derived fromperipheral blood and pre-activated through stimulation of the T-cellreceptor and CD28, proliferate in culture in response to the cytokineInterleukin-2 (IL-2). This proliferative response is dependent on theactivation of JAK1 and JAK3 tyrosine kinases, which phosphorylate andactivate the transcription factor Stat-5. The primary human T-cells areincubated with the 2,4-pyrimidinediamine compounds in the presence ofIL-2 for 72 hours, and at the assay endpoint intracellular ATPconcentrations are measured to assess cell viability. A reduction incell proliferation compared to control conditions is indicative ofinhibition of the JAK kinase pathway.

The activity of the compounds described herein can additionally becharacterized by assaying the effect of the 2,4-pyrimidinediaminecompounds described herein on A549 lung epithelial cells and U937 cells.A549 lung epithelial cells and U937 cells up-regulate ICAM-1 (CD54)surface expression in response to a variety of different stimuli.Therefore, using ICAM-1 expression as readout, test compound effects ondifferent signaling pathways can be assessed in the same cell type.Stimulation with IL-1β through the IL-1β receptor activates theTRAF6/NFκB pathway resulting in up-regulation of ICAM-1. IFNγ inducesICAM-1 up-regulation through activation of the JAK1/JAK2 pathway. Theup-regulation of ICAM-1 can be quantified by flow cytometry across acompound dose curve and EC₅₀ values are calculated.

Active compounds as described herein generally inhibit the JAK kinasepathway with an IC₅₀ in the range of about 1 mM or less, as measured inthe assays described herein. Of course, skilled artisans will appreciatethat compounds which exhibit lower IC₅₀s, (on the order, for example, of100 μM, 75 μM, 50 μM, 40 μM, 30 μM, 20 μM, 15 μM, 10 μM, 5 μM, 1 μM, 500nM, 100 nM, 10 nM, 1 nM, or even lower) can be particularly useful intherapeutic applications. In instances where activity specific to aparticular cell type is desired, the compound can be assayed foractivity with the desired cell type and counter-screened for a lack ofactivity against other cell types. The desired degree of “inactivity” insuch counter screens, or the desired ratio of activity vs. inactivity,may vary for different situations and can be selected by the user.

The 2,4-pyrimidinediamine active compounds also typically inhibit IL-4stimulated expression of CD23 in B-cells with an IC₅₀ in the range ofabout 20 μM or less, typically in the range of about 10 μM, 1 μM, 500nM, 100 nM, 10 nM, 1 nM, or even lower. A suitable assay that can beused is the assay described in Example 2, “Assay for Ramos B-Cell LineStimulated with IL-4.” In certain embodiments, the active2,4-pyrimidinediamine compounds have an IC₅₀ of less than or equal to 5μM, greater than 5 μM but less than 20 μM, greater than 20 μM, orgreater than 20 μM but less than 50 μM in the assay described in Example2.

Additionally, the 2,4-pyrimidinediamine active compounds typicallyinhibit an activity of human primary T-cells with an IC₅₀ in the rangeof about 20 μM or less, typically in the range of about 10 μM, 1 μM, 500nM, 100 nM, 10 nM, 1 nM, or even lower. The IC₅₀ against human primaryT-cells can be determined in a standard in vitro assay with isolatedhuman primary T-cells. A suitable assay that can be used is the assaydescribed above, “Primary Human T-cell Proliferation Assay Stimulatedwith IL-2.” In some embodiments, the active 2,4-pyrimidinediaminecompounds have an IC₅₀ of less than or equal to 5 μM, greater than 5 μMbut less than 20 μM, greater than 20 μM, or greater than 20 μM but lessthan 50 μM in the assay described above.

The 2,4-pyrimidinediamine active compounds also typically inhibitexpression of ICAM1 (CD54) induced by IFNγ, exposure in U937 or A549cells with an IC₅₀ in the range of about 20 μM or less, typically in therange of about 10 μM, 1 μM, 500 nM, 100 nM, 10 nM, 1 nM, or even lower.The IC₅₀ against expression of ICAM (CD54) in IFNγ stimulated cells canbe determined in a functional cellular assay with an isolated A549 orU937 cell line. The active 2,4-pyrimidinediamine compounds typicallyhave an IC₅₀ of less than or equal to 20 μM, greater than 20 μM, orgreater than 20 μM but less than 50 μM in the assay.

E. Pharmaceutical Compositions of the Invention

Pharmaceutical compositions including the 2,4-pyrimidinediaminecompounds described herein (or prodrugs thereof) can be manufactured bymeans of conventional mixing, dissolving, granulating, dragee-making,levigating, emulsifying, encapsulating, entrapping or lyophilizationprocesses. The compositions can be formulated in conventional mannerusing one or more physiologically acceptable carriers, diluents,excipients or auxiliaries which facilitate processing of the activecompounds into preparations which can be used pharmaceutically.

The 2,4-pyrimidinediamine compound or prodrug can be formulated in thepharmaceutical compositions per se, or in the form of a hydrate,solvate, N-oxide or pharmaceutically acceptable salt, as describedherein. Typically, such salts are more soluble in aqueous solutions thanthe corresponding free acids and bases, but salts having lowersolubility than the corresponding free acids and bases may also beformed.

In one embodiment, this invention provides a pharmaceutical formulationincluding a compound selected from the compounds of this invention, asdescribed above.

The compounds can be provided in a variety of formulations and dosages.The compounds can be provided in a pharmaceutically acceptable formincluding, where the compound or prodrug can be formulated in thepharmaceutical compositions per se, or in the form of a hydrate,solvate, N-oxide or pharmaceutically acceptable salt, as describedherein. Typically, such salts are more soluble in aqueous solutions thanthe corresponding free acids and bases, but salts having lowersolubility than the corresponding free acids and bases may also beformed. It is to be understood that reference to the compound,2,4-pyrimidinediamine compound, or “active” in discussions offormulations is also intended to include, where appropriate as known tothose of skill in the art, formulation of the prodrugs of the2,4-pyrimidinediamine compounds.

In one embodiment, the compounds are provided as non-toxicpharmaceutically acceptable salts, as noted previously. Suitablepharmaceutically acceptable salts of the compounds of this inventioninclude acid addition salts such as those formed with hydrochloric acid,fumaric acid, p-toluenesulphonic acid, maleic acid, succinic acid,acetic acid, citric acid, tartaric acid, carbonic acid or phosphoricacid. Salts of amine groups may also include quaternary ammonium saltsin which the amino nitrogen atom carries a suitable organic group suchas an alkyl, alkenyl, alkynyl or aralkyl moiety. Furthermore, where thecompounds of the invention carry an acidic moiety, suitablepharmaceutically acceptable salts thereof may include metal salts suchas alkali metal salts, e.g. sodium or potassium salts; and alkalineearth metal salts, e.g. calcium or magnesium salts.

The pharmaceutically acceptable salts of the present invention can beformed by conventional means, such as by reacting the free base form ofthe product with one or more equivalents of the appropriate acid in asolvent or medium in which the salt is insoluble, or in a solvent suchas water which is removed in vacuo or by freeze drying or by exchangingthe anions of an existing salt for another anion on a suitable ionexchange resin.

The present invention includes within its scope solvates of the2,4-pyrimidinediamine compounds and salts thereof, for example,hydrates.

The 2,4-pyrimidinediamine compounds may have one or more asymmetriccenters, and may accordingly exist both as enantiomers and asdiastereomers. It is to be understood that all such isomers and mixturesthereof are encompassed within the scope of the present invention.

The 2,4-pyrimidinediamine compounds can be administered by oral,parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV,intracisternal injection or infusion, subcutaneous injection, orimplant), by inhalation spray, nasal, vaginal, rectal, sublingual,urethral (e.g., urethral suppository) or topical routes ofadministration (e.g., gel, ointment, cream, aerosol, etc.) and can beformulated, alone or together, in suitable dosage unit formulationscontaining conventional non-toxic pharmaceutically acceptable carriers,adjuvants, excipients and vehicles appropriate for each route ofadministration. In addition to the treatment of warm-blooded animalssuch as mice, rats, horses, cattle, sheep, dogs, cats, monkeys, etc.,the compounds of the invention can be effective in humans.

The pharmaceutical compositions for the administration of the2,4-pyrimidinediamine compounds may conveniently be presented in dosageunit form and can be prepared by any of the methods well known in theart of pharmacy. The pharmaceutical compositions can be, for example,prepared by uniformly and intimately bringing the active ingredient intoassociation with a liquid carrier or a finely divided solid carrier orboth, and then, if necessary, shaping the product into the desiredformulation. In the pharmaceutical composition the active objectcompound is included in an amount sufficient to produce the desiredtherapeutic effect. For example, pharmaceutical compositions of theinvention may take a form suitable for virtually any mode ofadministration, including, for example, topical, ocular, oral, buccal,systemic, nasal, injection, transdermal, rectal, vaginal, etc., or aform suitable for administration by inhalation or insufflation.

The 2,4-pyrimidinediamine compound(s) or prodrug(s) described herein, orcompositions thereof, will generally be used in an amount effective toachieve the intended result, for example in an amount effective to treator prevent the particular condition being treated. The compound(s) canbe administered therapeutically to achieve therapeutic benefit orprophylactically to achieve prophylactic benefit. By therapeutic benefitis meant eradication or amelioration of the underlying disorder beingtreated and/or eradication or amelioration of one or more of thesymptoms associated with the underlying disorder such that the patientreports an improvement in feeling or condition, notwithstanding that thepatient may still be afflicted with the underlying disorder. Forexample, administration of a compound to a patient suffering from anallergy provides therapeutic benefit not only when the underlyingallergic response is eradicated or ameliorated, but also when thepatient reports a decrease in the severity or duration of the symptomsassociated with the allergy following exposure to the allergen. Asanother example, therapeutic benefit in the context of asthma includesan improvement in respiration following the onset of an asthmaticattack, or a reduction in the frequency or severity of asthmaticepisodes. As another specific example, therapeutic benefit in thecontext of transplantation rejection includes the ability to alleviatean acute rejection episode, such as for example, HVGR or GVHR, or theability to prolong the time period between onset of acute rejectionepisodes and/or onset of chronic rejection. Therapeutic benefit alsoincludes halting or slowing the progression of the disease, regardlessof whether improvement is realized.

The amount of compound administered will depend upon a variety offactors, including, for example, the particular condition being treated,the mode of administration, the severity of the condition being treatedand the age and weight of the patient, the bioavailability of theparticular active compound, etc. Determination of an effective dosage iswell within the capabilities of those skilled in the art.

As known by those of skill in the art, the preferred dosage of2,4-pyrimidinediamine compounds will also depend on the age, weight,general health and severity of the condition of the individual beingtreated. Dosage may also need to be tailored to the sex of theindividual and/or where administered by inhalation, the lung capacity ofthe individual. Dosage may also be tailored to individuals sufferingfrom more than one condition or those individuals who have additionalconditions which affect lung capacity and the ability to breathenormally, for example, emphysema, bronchitis, pneumonia, respiratoryinfections, etc. Dosage, and frequency of administration of thecompounds or prodrugs thereof, will also depend on whether the compoundsare formulated for treatment of acute episodes of a condition or for theprophylactic treatment of a disorder. For example, acute episodes ofallergic conditions, including allergy-related asthma, transplantrejection, etc. A skilled practitioner will be able to determine theoptimal dose for a particular individual.

For prophylactic administration, the compound can be administered to apatient at risk of developing one of the previously describedconditions. For example, if it is unknown whether a patient is allergicto a particular drug, the compound can be administered prior toadministration of the drug to avoid or ameliorate an allergic responseto the drug. Alternatively, prophylactic administration can be appliedto avoid the onset of symptoms in a patient diagnosed with theunderlying disorder. For example, a compound can be administered to anallergy sufferer prior to expected exposure to the allergen. Compoundsmay also be administered prophylactically to healthy individuals who arerepeatedly exposed to agents known to one of the above-describedmaladies to prevent the onset of the disorder. For example, a compoundcan be administered to a healthy individual who is repeatedly exposed toan allergen known to induce allergies, such as latex, in an effort toprevent the individual from developing an allergy. Alternatively, acompound can be administered to a patient suffering from asthma prior topartaking in activities which trigger asthma attacks to lessen theseverity of, or avoid altogether, an asthmatic episode.

In the context of transplant rejection, the compound can be administeredwhile the patient is not having an acute rejection reaction to avoid theonset of rejection and/or prior to the appearance of clinicalindications of chronic rejection. The compound can be administeredsystemically to the patient as well as administered to the tissue ororgan prior to transplanting the tissue or organ in the patient.

Also provided are kits for administration of the 2,4-substitutedpyrimidinediamine, prodrug thereof or pharmaceutical formulationsincluding the compound, that may include a dosage amount of at least one2,4-pyrimidinediamine or a composition including at least one2,4-pyrimidinediamine as disclosed herein. Kits may further includesuitable packaging and/or instructions for use of the compound. Kits mayalso include a means for the delivery of the at least one2,4-pyrimidinediamine or compositions including at least one2,4-substituted pyrimidinediamine, such as an inhaler, spray dispenser(e.g. nasal spray), syringe for injection or pressure pack for capsules,tables, suppositories, or other device as described herein.

It will be appreciated by one of skill in the art that the embodimentssummarized above may be used together in any suitable combination togenerate additional embodiments not expressly recited above, and thatsuch embodiments are considered to be part of the present invention.

F. General Synthesis of the Compounds of the Invention

The 2,4-pyrimidinediamine compounds and prodrugs of the invention can besynthesized via a variety of different synthetic routes usingcommercially available starting materials and/or starting materialsprepared by conventional synthetic methods. Suitable exemplary methodsthat can be routinely adapted to synthesize the 2,4-pyrimidinediaminecompounds and prodrugs of the invention are found in U.S. Pat. No.5,958,935, the disclosure of which is incorporated herein by reference.Specific examples describing the synthesis of numerous2,4-pyrimidinediamine compounds and prodrugs, as well as intermediatesthereof, are described in copending U.S. application Ser. No.10/355,543, filed Jan. 31, 2003 (US2004/0029902A1), the contents ofwhich are incorporated herein by reference. Suitable exemplary methodsthat can be routinely used and/or adapted to synthesize active2,4-pyrimidinediamine compounds can also be found in internationalapplication Serial No. PCT/US03/03022 filed Jan. 31, 2003 (WO03/063794), U.S. application Ser. No. 10/631,029 filed Jul. 29, 2003,international application Serial No. PCT/US03/24087 (WO2004/014382),U.S. application Ser. No. 10/903,263 filed Jul. 30, 2004, andinternational application Serial No. PCT/US2004/24716 (WO005/016893),the disclosures of which are incorporated herein by reference. All ofthe compounds described herein (including prodrugs) can be prepared byroutine adaptation of these methods.

Specific exemplary synthetic methods for the 2,4-pyrimidinediaminesdescribed herein are also described in Example 1 below. Those of skillin the art will also be able to readily adapt these examples for thesynthesis of additional 2,4-pyrimidinediamines as described herein.

A variety of exemplary synthetic routes that can be used to synthesizethe 2,4-pyrimidinediamine compounds of the invention are described inSchemes (I)-(VII), below. These methods can be routinely adapted tosynthesize the 2,4-pyrimidinediamine compounds and prodrugs describedherein.

In one exemplary embodiment, the compounds can be synthesized fromsubstituted or unsubstituted uracils as illustrated in Scheme (I),below:

In Scheme (I), R¹, (R²)_(p), (R³)_(q), R⁴, R⁵, and X, are as definedherein, and Q is —YN(R⁶)S(O)₂N(R⁷)R⁸, wherein R⁶, R⁷ and R⁸ are asdefined herein. According to Scheme (I), uracil A-1 is dihalogenated atthe 2- and 4-positions using a standard halogenating agent such as POCl₃(or other standard halogenating agent) under standard conditions toyield 2,4-dichloropyrimidine A-2. Depending upon the X substituent, inpyrimidinediamine A-2, the chloride at the C4 position is more reactivetowards nucleophiles than the chloride at the C2 position. Thisdifferential reactivity can be exploited to synthesize2,4-pyrimidinediamines I by reacting 2,4-dichloropyrimidine A-2 firstwith one equivalent of amine A-3, yielding4N-substituted-2-chloro-4-pyrimidineamine A-4, and then with amine A-5,yielding a 2,4-pyrimidinediamine derivative A-6, where N4 nitrogen canbe selectively alkylated to give compounds of formula I.

Typically, the C4 halide is more reactive towards nucleophiles, asillustrated in Scheme (I). However, as will be recognized by skilledartisans, the identity of the X substituent may alter this reactivity.For example, when X is trifluoromethyl, a 50:50 mixture of4N-substituted-4-pyrimidineamine A-4 and the corresponding2N-substituted-2-pyrimidineamine is obtained. The regioselectivity ofthe reaction can also be controlled by adjusting the solvent and othersynthetic conditions (such as temperature), as is well-known in the art.

The reactions depicted in Scheme (I) may proceed more quickly when thereaction mixtures are heated via microwave. When heating in thisfashion, the following conditions can be used: heat to 175° C. inethanol for 5-20 min in a Smith Reactor (Personal Chemistry, Uppsala,Sweden) in a sealed tube (at 20 bar pressure).

The uracil A-1 starting materials can be purchased from commercialsources or prepared using standard techniques of organic chemistry.Commercially available uracils that can be used as starting materials inScheme (I) include, by way of example and not limitation, uracil(Aldrich #13, 078-8; CAS Registry 66-22-8); 5-bromouracil (Aldrich #85,247-3; CAS Registry 51-20-7; 5-fluorouracil (Aldrich #85, 847-1; CASRegistry 51-21-8); 5-iodouracil (Aldrich #85, 785-8; CAS Registry696-07-1); 5-nitrouracil (Aldrich #85, 276-7; CAS Registry 611-08-5);and 5-(trifluoromethyl)-uracil (Aldrich #22, 327-1; CAS Registry54-20-6). Additional 5-substituted uracils are available from GeneralIntermediates of Canada, Inc., Edmonton, Calif., and/or Interchim,Cedex, France, or can be prepared using standard techniques. Myriadtextbook references teaching suitable synthetic methods are providedinfra.

Amines A-3 and A-5 can be purchased from commercial sources or,alternatively, can be synthesized using standard techniques. Forexample, suitable amines can be synthesized from nitro precursors usingstandard chemistry. Specific exemplary reactions are provided in theExamples section. See also Vogel, 1989, Practical Organic Chemistry,Addison Wesley Longman, Ltd. and John Wiley & Sons, Inc.

Skilled artisans will recognize that in some instances, amines A-3 andA-5 and/or substituent X on uracil A-1 may include functional groupsthat require protection during synthesis. The exact identity of anyprotecting group(s) used will depend upon the identity of the functionalgroup being protected, and will be apparent to those of skill in theart. Guidance for selecting appropriate protecting groups, as well assynthetic strategies for their attachment and removal, can be found, forexample, in Greene & Wuts, Protective Groups in Organic Synthesis, 3dEdition, John Wiley & Sons, Inc., New York (1999) and the referencescited therein (hereinafter “Greene & Wuts”).

Thus, “protecting group” refers to a group of atoms that, when attachedto a reactive functional group in a molecule, mask, reduce or preventthe reactivity of the functional group. Typically, a protecting groupcan be selectively removed as desired during the course of a synthesis.Examples of protecting groups can be found in Greene and Wuts, asmentioned above, and, additionally, in Harrison et al., Compendium ofSynthetic Organic Methods, Vols. 1-8, 1971-1996, John Wiley & Sons, NY.Representative amino protecting groups include, but are not limited to,formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl (“CBZ”),tert-butoxycarbonyl (“Boc”), trimethylsilyl (“TMS”),2-trimethylsilyl-ethanesulfonyl (“TES”), trityl and substituted tritylgroups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (“FMOC”),nitro-veratryloxycarbonyl (“NVOC”), and the like. Representativehydroxyl protecting groups include, but are not limited to, those wherethe hydroxyl group is either acylated to form acetate and benzoateesters or alkylated to form benzyl and trityl ethers, as well as alkylethers, tetrahydropyranyl ethers, trialkylsilyl ethers (e.g., TMS orTIPPS groups), aryl silyl ethers (e.g., triphenylsilyl ether), mixedalkyl and aryl substituted silyl ethers, and allyl ethers.

A specific embodiment of Scheme (I) utilizing 5-fluorouracil (Aldrich#32, 937-1) as a starting material is illustrated in Scheme (Ia), below:

In Scheme (Ia), (R²)_(p), (R³)_(q), R⁴, R⁵, and Q are as previouslydefined for Scheme (I). Asymmetric2N,4N-disubstituted-5-fluoro-2,4-pyrimidinediamine A-11 can be obtainedby reacting 2,4-dichloro-5-fluoropyrimidine A-9 with one equivalent ofamine A-3 (to yield 2-chloro-N4-substituted-5-fluoro-4-pyrimidineamineA-10) followed by one or more equivalents of amine A-5.

Myriad references teaching methods useful for synthesizing pyrimidinesgenerally, as well as starting materials described above, are known inthe art. For specific guidance, the reader is referred to Brown, D. J.,“The Pyrimidines”, in The Chemistry of Heterocyclyl Compounds, Volume 16(Weissberger, A., Ed.), 1962, Interscience Publishers, (A Division ofJohn Wiley & Sons), New York (“Brown I”); Brown, D. J., “ThePyrimidines”, in The Chemistry of Heterocyclyl Compounds, Volume 16,Supplement I (Weissberger, A. and Taylor, E. C., Ed.), 1970,Wiley-Interscience, (A Division of John Wiley & Sons), New York (BrownII″); Brown, D. J., “The Pyrimidines”, in The Chemistry of HeterocyclylCompounds, Volume 16, Supplement II (Weissberger, A. and Taylor, E. C.,Ed.), 1985, An Interscience Publication (John Wiley & Sons), New York(“Brown III”); Brown, D. J., “The Pyrimidines” in The Chemistry ofHeterocyclyl Compounds, Volume 52 (Weissberger, A. and Taylor, E. C.,Ed.), 1994, John Wiley & Sons, Inc., New York, pp. 1-1509 (Brown IV″);Kenner, G. W. and Todd, A., in Heterocyclyl Compounds, Volume 6,(Elderfield, R. C., Ed.), 1957, John Wiley, New York, Chapter 7(pyrimidines); Paquette, L. A., Principles of Modern HeterocyclylChemistry, 1968, W. A. Benjamin, Inc., New York, pp. 1-401 (uracilsynthesis pp. 313, 315; pyrimidinediamine synthesis pp. 313-316; aminopyrimidinediamine synthesis pp. 315); Joule, J. A., Mills, K. and Smith,G. F., Heterocyclyl Chemistry, 3^(rd) Edition, 1995, Chapman and Hall,London, UK, pp. 1-516; Vorbrüggen, H. and Ruh-Pohlenz, C., Handbook ofNucleoside Synthesis, John Wiley & Sons, New York, 2001, pp. 1-631(protection of pyrimidines by acylation pp. 90-91; silylation ofpyrimidines pp. 91-93); Joule, J. A., Mills, K. and Smith, G. F.,Heterocyclyl Chemistry, 4^(th) Edition, 2000, Blackwell Science, Ltd,Oxford, UK, pp. 1-589; and Comprehensive Organic Synthesis, Volumes 1-9(Trost, B. M. and Fleming, I., Ed.), 1991, Pergamon Press, Oxford, UK.

EXAMPLES

The invention is further understood by reference to the followingexamples, which are intended to be purely exemplary of the invention.The present invention is not limited in scope by the exemplifiedembodiments, which are intended as illustrations of single aspects ofthe invention only. Any methods that are functionally equivalent arewithin the scope of the invention. Various modifications of theinvention in addition to those described herein will become apparent tothose skilled in the art from the foregoing description and accompanyingfigures. Such modifications fall within the scope of the appendedclaims.

In the examples below as well as throughout the application, thefollowing abbreviations have the following meanings. If not defined, theterms have their generally accepted meanings.

mL = milliliter s = singlet d = doublet t = triplet q = quartet m =multiplet dd = doublet of doublets br = broad nM = nanomolar μg =microgram ng = nanogram MS = mass spectrum or mass spectrometry LC =liquid chromatography DMSO = dimethylsulfoxide μL = microliter mM =millimolar rpm = revolutions per minute LAH = lithium aluminum hydrideHEPES = N-(2-hydroxyethyl)-piperazine-N′-2-ethanesulfonic acid EGTA =Ethylene glycol-bis(beta-aminoethyl ether)-N,N,N′,N′-tetraacetic acidBRIJ-35 = polyoxyethyleneglycol dodecyl ether detergent

Example 1N2-(3-aminosulfonyl-4-methyl)phenyl-N4-(3-ethylaminosulfonyl-aminomethyl)phenyl-5-methyl-2,4-pyrimidinediamine

Tert-butyl N-(3-nitrobenzylsulfamoyl)carbamate

3-Nitrobenzylamine HCl salt (1 g, 5.3 mmol) and 4-Pyridinaminium,1-[[[(1,1-dimethylethoxy)carbonyl]amino]sulfonyl]-1,4-dihydro-N,N-dimethyl-,inner salt (2.4 g) were dissolved in dichloromethane (20 mL) andtriethylamine (2.2 mL). The solution was stirred at ambient temperaturefor three days. The reaction mixture was diluted with 1N HCl aq. (100mL) and extracted with ethyl acetate (3×100 mL). The organic solutionswere evaporated to give tert-butyl N-(3-nitrobenzylsulfamoyl)carbamatewhich was not isolated. MS (m/e): 330.11 (MH−).

N-(3-nitrobenzyl)-N′-ethylsulfamide

Crude tert-butyl N-(3-nitrobenzylsulfamoyl)carbamate, triphenylphosphine(2 g) and diisopropyl azadicarboxylate (DIAD, 1.6 g) were dissolved inTHF (20 mL) and ethanol (0.47 mL). The solution was stirred at ambienttemperature overnight. The reaction solution was evaporated to dryness.The residue was dissolved in methanol (10 mL) and treated with 4.0 M HClin dioxane (10 mL). It was stirred at rt overnight and then evaporated.The residue was diluted with ethyl acetate (100 mL) and washed withNaHCO3 aq. (2×100 mL). The organic solution was evaporated and purifiedby flash column chromatography (EtOAc/hexane=1/2, 1/1) to giveN-(3-nitrobenzyl)-N′-ethylsulfamide. ¹H NMR (DMSO-d₆): δ 1.02 (t, J=7.2Hz, 3H), 2.82 (p, J=6.9 Hz, 2H), 4.14 (d, J=6.6 Hz, 2H), 6.93 (t, J=5.4Hz, 1H), 7.54 (t, J=6.3 Hz, 1H), 7.61 (t, J=7.8 Hz, 1H), 7.77 (d, J=7.2Hz, 1H), 8.10 (d, J=8.1 Hz, 1H), 8.20 (s, 1H); LCMS: purity: 1.2 g,88.4%; MS (m/e): 260.16 (MH+).

N-(3-nitrobenzyl)-N′-ethylsulfamide was dissolved in methanol (100 mL)and to the solution was added 10% Pd—C. The reaction mixture was reactedunder hydrogen atmosphere (˜40 psi) for 1 h. The catalyst was filteredoff over celite and washed with methanol. The filtrate was evaporated togive N-(3-aminobenzyl)-N′-ethylsulfamide. ¹H NMR (DMSO-d₆): δ 1.03 (t,J=7.2 Hz, 3H), 2.83 (p, J=6.9 Hz, 2H), 3.81 (d, J=6.3 Hz, 2H), 5.02 (s,2H), 6.41 (t, J=7.8 Hz, 2H), 6.51 (s, 1H), 6.76 (t, J=5.7 Hz, 1H), 6.91(t, J=7.5 Hz, 1H), 7.16 (t, J=6.3 Hz, 1H).

2-Chloro-N4-(3-ethylaminosulfonylaminomethyl)phenyl-5-methyl-4-pyrimidineamine

N-(3-Aminobenzyl)-N′-ethylsulfamide (1.2 g, 4.7 mmol,2,4-dichloro-5-methylpyrimidine (1.5 g, 9.2 mmol) and NaHCO3 (1 g) weredissolved in methanol (10 mL) and water (1 mL). The reaction solutionwas stirred at 60° C. overnight. The reaction mixture was diluted with1N HCl aq. (100 mL) and extracted with ethyl acetate (2×100 mL). Theorganic solutions were evaporated and purified by flash columnchromatography (EtOAc/hexanes=1/4, 1/2, 1/1, EtOAc) to give2-chloro-N4-(3-ethylaminosulfonylaminomethyl)phenyl-5-methyl-4-pyrimidineamine.LCMS: purity: 91.88%; MS (m/e): 356.46 (MH⁺).

N2-(3-Aminosulfonyl-4-methyl)phenyl-N4-(3-ethylaminosulfonylaminomethyl)phenyl-5-methyl-2,4-pyrimidinediamine

2-Chloro-N4-(3-ethylaminosulfonylaminomethyl)phenyl-5-methyl-4-pyrimidineamine(50 mg) and (3-aminosulfonyl-4-methyl)aniline (50 mg) were suspended inisopropanol (1 mL) and TFA (5 drops). The solution was heated at 100° C.overnight, then cooled to room temperature. The solution was evaporatedand purified by HPLC to giveN2-(3-aminosulfonyl-4-methyl)phenyl-N4-(3-ethylaminosulfonylaminomethyl)phenyl-5-methyl-2,4-pyrimidinediamine.¹H NMR (DMSO-d₆): δ 1.02 (t, J=7.2 Hz, 3H), 2.16 (s, 3H), 2.52 (s, 3H),2.83 (m, J=6.3 Hz, 2H), 4.00 (d, J=6.3 Hz, 2H), 6.84 (t, 1H), 7.19 (m,2H), 7.34 (m, 4H), 7.43 (s, 1H), 7.49 (d, 1H), 7.74 (m, 2H), 7.86 (s,1H), 9.58 (br, 1H), 10.11 (br, 1H); LCMS: purity: 97.72%; MS (m/e):506.25 (MH⁺).

The following examples were made using methods analogous to the exampleabove:

I-1:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N,N-dimethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 513 (MH⁺). ¹H NMR (DMSO-d6):

9.98 (s, 1H), 9.29 (s, 1H), 8.23 (s, 1H), 7.93-7.89 (m, 2H), 7.71 (t,1H, J=6.4 Hz), 7.59 (d, 2H, J=8.2 Hz), 7.40-7.30 (m, 6H), 4.09 (s, 2H),2.63 (s, H).

I-2:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N,N-dimethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 513 (MH⁺). ¹H NMR (DMSO-d6):

10.91 (s, 1H), 9.18 (s, 1H), 8.23 (s, 1H), 7.75-7.72 (app m, 3H, J=8.8Hz), 7.59 (m, 4H), 7.34 (d, 2H, J=8.5 Hz), 7.15 (br s, 2H), 4.11 (m,2H), 2.64 (s, 6H).

I-3:N2-(3-Aminosulfonylphenyl)-N-4-[4-[(N,N-dimethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 492 (MH⁺). ¹H NMR (DMSO-d6):

10.63 (s, 1H), 9.77 (s, 1H), 7.93 (s, 1H), 7.82 (d, 1H, J=8.5 Hz), 7.72(t, 1H, J=6.7 Hz), 7.65 (s, 1H), 7.51-7.48 (m, 3H), 7.42-7.34 (m, 5H),4.12 (d, 2H, J=6.7 Hz), 2.64 (s, 6H), 2.16 (s, 3H).

I-4:N2-(4-Aminosulfonylphenyl)-N4-[4-[(N,N-dimethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 492 (MH⁺). ¹H NMR (DMSO-d6):

10.60 (s, 1H), 9.65 (s, 1H), 7.95 (s, 1H), 7.72 (t, 1H, J=6.2 Hz),7.66-7.51 (m, 6H), 7.38 (d, 2H, J=8.5 Hz), 7.24 (s, 2H), 4.14 (d, 2H,J=6.2 Hz), 2.64 (s, 6H), 2.17 (s, 3H).

I-5:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-ethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 513 (MN). ¹H NMR (DMSO-d6):

0.72 (s, 1H), 9.00 (s, 1H), 8.18 (s, 1H), 7.94 (s, 1H), 7.74 (br s, 2H),7.61 (d, 2H, J=8.5 Hz), 7.36-7.27 (m, 7H), 3.99 (br s, 2H), 2.85 (qt,2H, J=7.3 Hz), 1.02 (t, 3H, J=7.3 Hz).

I-6:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-ethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine

LCMS: Purity: 97%; MS (m/e): 513 (MH⁺). ¹H NMR (DMSO-d6):

0.78 (s, 1H), 9.05 (s, 1H), 8.20 (s, 1H), 7.76 (d, 2H, J=8.8 Hz), 7.60(d, 4H, J=8.8 Hz), 7.33 (app d, 3H, J=8.8 Hz), 6.73 (br s, 1H), 4.01 (s,2H), 2.85 (qt, 2H, J=7.3 Hz), 1.02 (t, 3H, J=7.3 Hz).

I-7:N2-(3-Aminosulfonylphenyl)-N4-[4-[(N-ethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine

LCMS: Purity: 97%; MS (m/e): 492 (MH⁺). ¹H NMR (DMSO-d6):

10.59 (s, 1H), 9.76 (s, 1H), 7.92 (s, 1H), 7.82 (d, 1H, J=8.2 Hz), 7.65(s, 1H), 7.49-7.42 (m, 3H), 7.38-7.33 (m, 6H), 6.89 (s, 1H), 4.01 (d,2H, J=5.2 Hz), 2.84 (app m, 2H), 2.16 (s, 3H), 1.03 (t, 3H, J=7.3 Hz).

I-8:N2-(4-Aminosulfonylphenyl)-N4-[4-[(N-ethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 492 (MH⁺). ¹H NMR (DMSO-d6):

10.69 (s, 1H), 9.77 (s, 1H), 7.95 (s, 1H), 7.65 (d, 2H, J=9.1 Hz), 7.5(d, 2H, J=9.1 Hz), 7.51 (d, 2H, J=8.8 Hz), 7.37 (app d, 3H, J=8.8 Hz),7.25 (s, 2H), 6.88 (s, 1H), 4.02 (d, 2H, J=5.2 Hz), 2.86 (app qt, 2H,J=7.3 Hz), 1.02 (t, 3H, J=7.3 Hz).

I-9:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine

LCMS: Purity: 97%; MS (m/e): 513 (MH⁺). ¹H NMR (DMSO-d6):

0.82 (s, 1H), 9.20 (s, 1H), 8.22 (s, 1H), 7.91 (m, 2H), 7.59-7.54 (m,2H), 7.39-7.03 (m, 5H), 7.16 (d, 1H, J=7.3 Hz), 4.00 (s, 2H), 2.82 (qt,2H, J=7.3 Hz), 1.02 (t, 3H, J=7.3 Hz).

I-10:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine

LCMS: Purity: 90%; MS (m/e): 513 (MH⁺). ¹H NMR (DMSO-d6):

9.83 (s, 1H), 9.19 (s, 1H), 8.22 (s, 1H), 7.73-7.70 (m, 2H), 7.58-7.51(m, 4H), 7.35-7.00 (m, 2H), 7.19-7.13 (m, 2H), 4.02 (s, 2H), 2.82 (qt,2H, J=7.3 Hz), 1.02 (m, 3H).

I-11:N2-(3-Aminosulfonylphenyl)-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 492 (MH⁺). ¹H NMR (DMSO-d6):

10.50 (s, 1H), 9.76 (s, 1H), 7.93 (s, 1H), 7.82 (d, 1H, J=8.5 Hz), 7.65(s, 1H), 7.50-7.48 (m, 2H), 7.42-7.33 (m, 6H), 7.21 (d, 1H, J=7.6 Hz),6.85-6.84 (m, 1H), 3.99 (d, 2H, J=6.2 Hz), 2.81-2.80 (m, 2H), 2.16 (s,3H), 1.02 (t, 3H, J=7.3 Hz).

I-12:N2-(4-Aminosulfonylphenyl)-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine

LCMS: Purity: 90%; MS (m/e): 492 (MH⁺). ¹H NMR (DMSO-d6):

0.41 (s, 1H), 8.51 (s, 1H), 7.93 (s, 1H), 7.81-7.78 (d, 2H, J=9.1 Hz),7.64 (d, 1H, J=8.2 Hz), 7.58-7.54 (m, 3H), 7.35-7.28 (m, 2H), 7.09-7.07(m, 3H), 6.82 (t, 1H, J=5.8 Hz), 4.01 (d, 2H, J=6.4 Hz), 2.82 (d qt, 2H,J=5.6 and 7.3 Hz), 2.12 (s, 3H), 1.01 (t, 3H, J=7.3 Hz).

I-13:N2-(3-Aminosulfonylphenyl)-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-5-fluoro-2,4-pyrimidinediamine

LCMS: Purity: 92%; MS (m/e): 496 (MH⁺). ¹H NMR (DMSO-d6):

0.78 (s, 1H), 8.19 (d, 1H, J=4.1 Hz), 8.00 (s, 1H), 7.93-7.90 (m, 1H),7.73 (d, 1H, J=7.6 Hz), 7.64 (s, 1H), 7.41-7.28 (m, 5H), 7.09 (d, 1H,J=6.7 Hz), 3.99 (s, 2H), 2.83 (app qt, 2H, J=7.3 Hz), 1.02 (t, 3H, J=7.3Hz).

I-14:N2-(4-Aminosulfonylphenyl)-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-5-fluoro-2,4-pyrimidinediamine

LCMS: Purity: 97%; MS (m/e): 496 (MH⁺). ¹H NMR (DMSO-d6):

9.71 (s, 1H), 9.67 (s, 1H), 8.18 (d, 1H, J=3.8 Hz), 7.80-7.71 (m, 3H),7.62 (app d, 3H, J=8.8 Hz), 7.35-7.29 (m, 1H), 7.13-7.01 (m, 3H), 4.00(s, 2H), 2.83 (qt, 2H, J=7.3 Hz), 1.02 (t, 3H, J=7.3 Hz).

I-15:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-ethylsulfamoylamino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 527 (MN). ¹H NMR (DMSO-d6):

0.49 (s, 1H), 8.73 (s, 1H), 7.86 (s, 1H), 7.75 (d, 1H, J=8.2 Hz), 7.34(t, 1H, J=6.7 Hz), 7.26-7.12 (m, 7H), 6.89 (t, 1H, J=5.6 Hz), 4.02 (d,2H, J=6.4 Hz), 2.88 (d qt, 2H, J=5.6 and 7.3 Hz), 2.14 (s, 3H), 1.06 (t,3H, J=7.3 Hz).

I-16:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-ethylsulfamoylamino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 527 (MH⁺). ¹H NMR (DMSO-d6):

9.58 (s, 1H), 8.80 (s, 1H0, 7.59 (d, 2H, J=8.5 Hz), 7.44 (d, 2H, J=8.5Hz), 7.34 (t, 1H, J=5.4 Hz), 7.27-7.20 (m, 3H), 7.05 (s, 2H), 6.87 (t,1H, J=6.2 Hz), 4.03 (d, 2H, J=6.4 Hz), 2.86 (m, 2H, J=7.0 Hz), 2.15 (s,3H), 11.05 (t, 3H, J=7.3 Hz).

I-17:N2-(4-Aminosulfonyl-3,5-dimethylphenyl)-5-chloro-N4-[4-[(N-ethylsulfamoylamino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 555 (MH⁺). ¹H NMR (DMSO-d6):

9.38 (s, 1H), 8.77 (s, 1H), 7.33 (t, 1H, J=5.2 Hz), 7.25-7.19 (m, 5H),6.95 (s, 2H), 6.89 (t, 1H, J=5.8 Hz), 3.98 (d, 2H, J=6.4 Hz), 2.87 (m,2H), 2.27 (s, 6H), 2.14 (s, 3H), 1.05 (t, 2H, J=7.3 Hz).

I-18:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 525 (MH⁺). ¹H NMR (DMSO-d6):

9.61 (s, 1H), 8.86 (s, 1H), 8.15 9s, 1H), 7.98 (s, 2H), 7.62 (d, 2H,J=8.5 Hz), 7.46 (t, 1H, J=5.4 Hz), 7.33-7.26 (m, 7H), 4.02 (d, 2H, J=6.4Hz), 2.31 (m, 1H), 0.54-0.51 (m, 4H).

I-19:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 525 (MH⁺).

I-20:N2-(4-Aminosulfonyl-3,5-dimethylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoyl-amino)methyl]phenyl]-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 553 (MH⁺). ¹H NMR (DMSO-d6):

9.48 (s, 1H), 8.91 (s, 1H0, 8.18 (s, 1H), 7.76-7.42 (m, 3H), 7.46 (t,1H, J=5.4 Hz), 7.33-7.30 (m, 4H), 7.11 (s, 2H), 4.00 (d, 2H, J=6.4 Hz),2.47 (s, 6H), 2.31-2.28 (m, 1H), 0.54-0.52 (m, 4H).

I-21:N2-(3-Aminosulfonylphenyl)-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 504 (MN). ¹H NMR (DMSO-d6):

0.28 (s, 1H), 8.28 (s, 1H), 8.07 (s, 1H), 8.01 (d, 1H, J=8.5 Hz), 7.89(s, 1H), 7.68 (d, 2H, J=9.1 Hz), 7.43-7.22 (m, 8H), 4.00 (d, 2H, J=6.4Hz), 2.31-2.30 (m, 1H), 2.10 (s, 3H), 0.54-0.52 (m, 4H).

I-22:N2-(4-Aminosulfonylphenyl)-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 504 (MH⁺). ¹H NMR (DMSO-d6):

9.38 (s, 1H), 8.35 (s, 1H), 7.92 (s, 1H), 7.84 (d, 2H, J=8.8 Hz), 7.65(d, 2H, J=8.8 Hz), 7.62 (d, 2H, J=8.8 Hz), 7.42 (t, 1H, J=6.4 Hz),7.32-7.29 (m, 3H), 7.07 (s, 2H), 4.03 (d, 2H, J=6.4 Hz), 2.31-2.30 (m,1H), 2.11 (s, 3H), 0.54-0.52 (m, 4H).

I-23:N2-(4-Aminosulfonyl-3,5-dimethylphenyl)-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 532 (MN). ¹H NMR (DMSO-d6):

9.16 (s, 1H), 8.32 (s, 1H), 7.90 (s, 1H), 7.62 (d, 2H, J=8.8 Hz), 7.44(s, 2H), 7.38 (t, 1H, J=6.4 Hz), 7.30-7.28 (m, 2H), 6.97 (s, 2H), 4.00(d, 2H, J=6.4 Hz), 2.42 (s, 6H), 2.31-2.30 (m, 1H), 2.10 (s, 3H),0.54-0.50 (m, 4H).

I-24:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 539 (MH⁺). ¹H NMR (DMSO-d6):

9.49 (s, 1H), 8.73 (s, 1H), 8.09 (s, 1H), 7.86 (s, 1H), 7.75 (d, 1H,J=8.5 Hz), 7.51 (t, 1H, J=6.7 Hz), 7.33 (s, 1H), 7.25-7.12 (m, 7H), 4.04(d, 2H, J=6.4 Hz), 2.36-2.31 (m, 1H), 2.14 (s, 3H), 0.59-0.54 (m, 4H).

I-25:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine

LCMS: Purity: 99%; MS (m/e): 539 (MH⁺). ¹H NMR (DMSO-d6):

9.56 (s, 1H), 8.80 (s, 1H), 8.13 (s, 1H), 7.58 (d, 2H, J=8.5 Hz), 7.49(t, 1H, J=6.7 Hz), 7.45 (d, 2H, J=8.8 Hz), 7.32-7.21 (m, 4H), 7.06 (s,1H), 4.06 (d, 2H, J=6.4 Hz), 2.32-2.31 (m, 1H), 2.15 (s, 3H), 0.58-0.51(m, 4H).

I-26:N2-(4-Aminosulfonyl-3,5-dimethylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoyl-amino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine

LCMS: Purity: 98%; MS (m/e): 566 (MH⁺). ¹H NMR (DMSO-d6):

9.38 (s, 1H), 8.77 (s, 1H), 8.10 (s, 1H), 7.46 (t, 1H, J=6.7 Hz), 7.34(s, 1H), 7.26-7.20 (m, 5H), 6.95 (s, 2H), 4.00 (d, 2H, J=6.4 Hz),2.31-2.300 (m, 1H), 2.27 (s, 6H), 2.14 (s, 3H), 0.55-0.51 (m, 4H).

I-27:N4-[3-(N-ethyl-N-propionyl)aminosulfonyl-(N-propionyl)aminomethyl]phenyl-5-methyl-N2-(3-propionylaminosulfonyl)phenyl-2,4-pyrimidinediamin

LCMS: purity: 90.67%; MS (m/e): 660.42 (MH+). ¹H NMR (DMSO-d₆): δ 0.88(t, J=7.5 Hz, 3H), 0.92 (t, J=7.5 Hz, 3H), 0.98 (t, J=7.2 Hz, 3H), 1.22(t, J=7.5 Hz, 3H), 2.13 (s, 3H), 2.20 (q, J=7.2 Hz, 2H), 2.56-2.64 (m,4H), 3.83 (q, J=7.8 Hz, 2H), 5.05 (s, 2H), 6.98 (d, J=8.1 Hz, 1H), 7.34(m, 3H), 7.65 (m, 2H), 7.92 (s, 1H), 8.08 (d, 1H), 8.19 (s, 1H), 8.43(s, 1H), 9.20 (s, 1H), 11.95 (s, 1H).

I-28:N4-[3-(N-ethyl-N-propionyl)aminosulfonyl-(N-propionyl)aminomethyl]phenyl-5-methyl-N2-(3-propionylaminosulfonyl)phenyl-2,4-pyrimidinediamineSodium Salt

LCMS: purity: 87.50%; MS (m/e): 660.42 (MH+). ¹H NMR (DMSO-d₆): δ 0.83(t, J=7.5 Hz, 3H), 0.92 (t, J=7.2 Hz, 3H), 0.99 (t, J=7.2 Hz, 3H), 1.23(t, J=6.9 Hz, 3H), 1.88 (q, J=7.5 Hz, 2H), 2.11 (s, 3H), 2.56-2.64 (m,4H), 3.82 (q, J=7.5 Hz, 2H), 5.05 (s, 2H), 6.94 (d, J=6.9 Hz, 1H), 7.11(t, J=7.5 Hz, 1H), 7.20 (d, J=7.2 Hz, 1H), 7.31 (t, J=7.8 Hz, 1H), 7.66(d, J=6.6 Hz, 1H), 7.77 (s, 1H), 7.83 (m, 2H), 7.89 (s, 1H), 8.31 (s,1H), 8.84 (s, 1H).

I-29:N2-(3-aminosulfonyl-4-methyl)phenyl-N4-(3-ethylaminosulfonylaminomethyl)phenyl-5-methyl-2,4-pyrimidinediamine

LCMS: purity: 97.72%; MS (m/e): 506.25 (MH+). ¹H NMR (DMSO-d₆): δ 1.02(t, J=7.2 Hz, 3H), 2.16 (s, 3H), 2.52 (s, 3H), 2.83 (m, J=6.3 Hz, 2H),4.00 (d, J=6.3 Hz, 2H), 6.84 (t, 1H), 7.19 (m, 2H), 7.34 (m, 4H), 7.43(s, 1H), 7.49 (d, 1H), 7.74 (m, 2H), 7.86 (s, 1H), 9.58 (br, 1H), 10.11(br, 1H).

I-30:N2-(3-aminosulfonyl)phenyl-N4-(3-N,N-dimethylaminosulfonylaminomethyl)-5-methyl-2,4-pyrimidinediamine

LCMS: purity: 96.46%; MS (m/e): 492.29 (MH+). ¹H NMR (DMSO-d₆): δ 2.15(s, 3H), 2.62 (s, 6H), 4.09 (d, J=6.3 Hz, 2H), 7.15 (d, 1H), 7.31 (s,2H), 7.35 (m, 2H), 7.42 (d, 1H), 7.49 (s, 1H), 7.55 (d, 1H), 7.69 (t,1H), 7.80 (s, 1H), 7.89 (m, 2H).

I-31:N2-(4-aminosulfonyl)phenyl-N4-(3-N,N-dimethylaminosulfonylaminomethyl)-5-methyl-2,4-pyrimidinediamine

LCMS: purity: 92.92%; MS (m/e): 492.30 (MH+). ¹H NMR (DMSO-d₆): δ 2.15(s, 3H), 2.62 (s, 6H), 4.10 (d, 2H), 7.18 (br, 3H), 7.36 (t, J=7.8 Hz,1H), 7.48 (s, 1H), 7.60 (m, 3H), 7.69 (m, 3H), 7.92 (s, 1H).

I-32:N2-(3-aminosulfonyl-4-methyl)phenyl-N4-(3-N,N-dimethylaminosulfonylamino-methyl)-5-methyl-2,4-pyrimidinediamine

LCMS: purity: 92.48%; MS (m/e): 506.29 (MH+). ¹H NMR (DMSO-d₆): δ 2.15(s, 3H), 2.62 (s, 6H), 4.09 (d, J=5.4 Hz, 2H), 7.16 (d, J=8.4 Hz, 2H),7.31 (m, 3H), 7.48 (s, 1H), 7.56 (d, 1H), 7.69 (m, 1H), 7.71 (m, 1H),7.80 (br, 1H), 7.85 (s, 1H).

II-1:N2-(3-aminosulfonyl-4-methyl)phenyl-N4-(2-N,N-dimethylaminosulfonyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-5-fluoro-2,4-pyrimidinediamine

C₂₂H₂₆FN₇O₄S₂. MS (ESI) m/z 535.92 (M+1)⁺. ¹H NMR (300 MHz, CD₃SOCD₃) δppm: 2.49 (s, 6H, 2XNCH₃), 2.74-2.82 (m, 7H, CH₃ and 2CH₂), 4.37 (s, 2H,CH₂), 7.12 (d, J=10.0 Hz, 1H, ArH), 7.20 (d, J=10.0 Hz, 1H, ArH), 7.24(s, 2H, NH₂), 7.51 (m, 1H, ArH), 7.71 (s, 1H, ArH), 7.93 (d, J=6.7 Hz,1H, ArH), 8.05 (s, 1H, ArH), 8.08 (d, J=6.7 Hz, 1H, ArH), 9.35 (s, 1H,NH), 9.45 (s, 1H, NH).

II-2:N2-(3-aminosulfonyl-4-methyl)phenyl-N4-(2-N,N-dimethylaminosulfonyl-2,3-dihydro-1H-isoindol-5-yl)-5-fluoro-2,4-pyrimidinediamine

C₂₁H₂₄FN₇O₄S₂. MS (ESI) m/z 550.94 (M+1)⁺. ¹H NMR (300 MHz, CD₃SOCD₃) δppm: 2.48 (s, 6H, 2XNCH₃), 2.81 (s, CH₃), 4.56 (s, 2H, CH₂), 7.18 (m,1H, ArH), 7.21 (m, 1H, ArH), 7.26 (s, 2H, NH₂), 7.59 (m, 1H, ArH), 7.83(m, 2H, 2ArH), 8.02 (m, 1H, ArH), 8.12 (d, J=6.7 Hz, 1H, ArH), 9.48 (s,1H, NH), 9.56 (s, 1H, NH).

II-3:N2-(4-aminosulfonyl)phenyl-N4-(2-N,N-dimethylaminosulfonyl-2,3-dihydro-1H-isoindol-5-yl)-5-methyl-2,4-pyrimidinediamine

LCMS: purity: 77.64%; MS (m/e): 504.26 (MH+). ¹H NMR (DMSO-d₆): δ 2.12(s, 3H), 2.81 (s, 6H), 4.60 (s, 4H), 7.08 (br, 2H), 7.28 (d, J=7.8 Hz,1H), 7.57 (d, J=8.7 Hz, 3H), 7.68 (s, 1H), 7.80 (d, J=8.1 Hz, 2H), 7.93(s, 1H), 8.45 (s, 1H), 9.38 (s, 1H).

II-4:N2-(3-aminosulfonyl)phenyl-N4-(2-N,N-dimethylaminosulfonyl-2,3-dihydro-1H-isoindol-5-yl)-5-methyl-2,4-pyrimidinediamine

LCMS: purity: 87.83%; MS (m/e): 504.53 (MH+). ¹H NMR (DMSO-d₆): δ 2.11(s, 3H), 2.81 (s, 6H), 4.58 (s, 4H), 7.24 (br, 2H), 7.29 (m, 3H), 7.57(d, J=9.6 Hz, 1H), 7.77 (s, 1H), 7.90 (s, 1H), 7.99 (d, J=7.2 Hz, 1H),8.05 (s, 1H), 8.37 (s, 1H), 9.27 (s, 1H).

II-5:N2-(3-aminosulfonyl-4-methyl)phenyl-N4-(2-N,N-dimethylaminosulfonyl-2,3-dihydro-1H-isoindol-5-yl)-5-methyl-2,4-pyrimidinediamine

LCMS: purity: 92.70%; MS (m/e): 518.35 (MH+). ¹H NMR (DMSO-d₆): δ 2.15(s, 3H), 2.53 (s, 3H), 2.80 (s, 6H), 4.53 (s, 2H), 4.59 (s, 2H), 7.17(d, J=9.0 Hz, 1H), 7.31 (m, 3H), 7.43 (d, 1H), 7.57 (s, 1H), 7.68 (d,1H), 7.79 (s, 1H), 7.86 (s, 1H).

Example 2 Cell Titer-Glo, Human Primary T Cell, IL2

Materials and Reagents: Dimethyl Sulfoxide (DMSO) (Sigma-Aldrich, Cat#D2650) was used as a control. The following reagents were used: FicollHypaque (Amersham Pharmacia, Cat# 17-1440-03), Anti-Human CD3 (BDPharmingen, Cat# 555336), Anti-Human CD28 (Immunotech, Cat# IM1376),Yssel's Media (Gemini Bio-products, Cat# 400-103), RPMI 1640 (Cellgro,Cat# 10-040-CM), Fetal Bovine Serum (JRH, Cat# 12106-500M), andAnti-Human IL-2 (R& D Systems, Cat# 202-IL).

Human primary T-cells were isolated and cultured according to thefollowing procedure. Whole blood was obtained from a healthy volunteer,mixed 1:1 with PBS, layered on to Ficoll Hypaque in 2:1 blood/PBS:ficollratio and centrifuged for 30 min at 4° C. at 1750 rpm. The lymphocytesat the serum: ficoll interface were recovered and washed twice with 5volumes of PBS. The cells were resuspended in Yssel's medium containing40 U/mL IL-2 and seeded into a flask pre-coated with 1 mg/mL anti-CD3and 5 mg/mL anti-CD28. The primary T-cells were stimulated for 3-4 days,then transferred to a fresh flask and maintained in RPMI with 10% FBSand 40 U/mL IL-2.

Primary T-cells were starved of IL-2 overnight and resuspended inYssel's medium at 2×10⁶ cells/mL. 50 μL of cell suspension containing 80U/mL IL-2 was added to each well of a flat bottom 96 well black plate.For the unstimulated control, IL-2 was omitted from the last column onthe plate. Compound was serially diluted in DMSO from 5 mM in 3-folddilutions, and then diluted 1:250 in Yssel's medium. 50 μL of 2×compound was added per well in duplicate and the cells were allowed toproliferate for 72 hours at 37° C.

Proliferation was measured using Cell Titer-Glo. The substrate wasthawed and allowed to come to room temperature. After mixing the CellTiter-Glo reagent and diluent together, 100 μL was added to each well.The plates were mixed on an orbital shaker for two minutes to inducelysis and incubated at room temperature for an additional ten minutes toallow the signal to equilibrate. The Luminescence was read on the WallacPlate Reader (Wallac Victor2 1420 Multilabel Counter).

Compounds tested in the assay described in Example 2, each having anIC₅₀ less than 0.5 μM, were I-1 through I-13, I-15, I-17 through I-21,and I-23 through I-24.

1. A compound of formula I:

wherein: p is 0, 1, 2, or 3; q is 0, 1, 2, or 3; X is hydrogen, alkyl,substituted alkyl, hydroxy, alkoxy, substituted alkoxy, amino,substituted amino, carboxyl, carboxyl ester, —C(O)N(R⁴)R⁵, cyano, halo,nitro, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkynyl or substituted cycloalkynyl; Y is a straight or branchedchain C₁₋₆ alkylene group, cycloalkylene or substituted cycloalkylene;R¹ is hydrogen, alkyl, substituted alkyl, cycloalkyl or substitutedcycloalkyl; each R² independently is alkyl, substituted alkyl, alkoxy,substituted alkoxy, amino, substituted amino, aryl, substituted aryl,aryloxy, substituted aryloxy, cyano, cycloalkyl, substituted cycloalkyl,cycloalkoxy, substituted cycloalkoxy, heteroaryl, substitutedheteroaryl, heteroaryloxy, substituted heteroaryloxy, heterocyclyl,substituted heterocyclyl, heterocyclyloxy, substituted heterocyclyloxy,aminoacyl, aminoacyloxy, carboxyl, carboxyl ester, carbonate ester,—C(O)N(R⁴)R⁵, nitro or halo; each R³ independently is alkyl, substitutedalkyl, alkoxy, substituted alkoxy, amino, substituted amino, aryl,substituted aryl, aryloxy, substituted aryloxy, cyano, cycloalkyl,substituted cycloalkyl, cycloalkoxy, substituted cycloalkoxy,heteroaryl, substituted heteroaryl, heteroaryloxy, substitutedheteroaryloxy, heterocyclyl, substituted heterocyclyl, heterocyclyloxy,substituted heterocyclyloxy, aminoacyl, aminoacyloxy, carboxyl, carboxylester, carbonate ester, nitro or halo; each R⁴ independently ishydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl,heterocyclyl, substituted heterocyclyl, carboxyl, carboxyl ester, aryl,substituted aryl, heteroaryl, substituted heteroaryl, acyl or M⁺ whereinM⁺ is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄, wherein each R⁹ independently is hydrogenor alkyl, and the nitrogen immediately adjacent to R⁵ is anionic; or R⁴and R⁵ together with the intervening atom or atoms bound thereto, form aheterocyclyl or substituted heterocyclyl group; each R⁵ independently ishydrogen, alkyl, substituted alkyl, amino, cycloalkyl, substitutedcycloalkyl, heterocyclyl, substituted heterocyclyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, carboxyl, carboxyl ester oracyl; R⁶ is hydrogen, alkyl, substituted alkyl, cycloalkyl, substitutedcycloalkyl, heterocyclyl, substituted heterocyclyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl or acyl; or R⁶ is a straight orbranched chain C₁₋₆ alkylene group, cycloalkylene or substitutedcycloalkylene, linking the nitrogen bearing R⁶ and the ring bearing Y R⁷is hydrogen, alkyl, substituted alkyl, cycloalkyl, substitutedcycloalkyl, heterocyclyl, substituted heterocyclyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl or acyl; R⁸ is hydrogen, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl or acyl; or R⁷ and R⁸ together, with nitrogen towhich they are bound, form a heterocyclyl or substituted heterocyclylgroup; wherein at least one of R⁷ and R⁸ is non-hydrogen; and each ofR⁶, R⁷, and R⁸ optionally are M⁺ wherein M⁺ is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄,wherein each R⁹ independently is hydrogen or alkyl, and the nitrogenbearing M⁺ is anionic.
 2. The compound of claim 1, wherein R¹ ishydrogen.
 3. The compound of claim 2, wherein X is alkyl, substitutedalkyl or halo.
 4. The compound of claim 1, wherein R⁴ is hydrogen and R⁵is hydrogen.
 5. The compound of claim 1, wherein Y is methylene.
 6. Thecompound of claim 1, wherein R⁶ is hydrogen, R⁷ is C₁₋₃ alkyl and R⁸ isC₁₋₃ alkyl.
 7. The compound of claim 1, wherein each of R² and R³,independently, is lower alkyl or lower alkoxy.
 8. The compound of claim1, according to Formula II:

wherein p is 0, 1, 2, or 3; q is 0, 1, 2, or 3; X is hydrogen, alkyl,substituted alkyl, hydroxy, alkoxy, substituted alkoxy, amino,substituted amino, carboxyl, carboxyl ester, —C(O)N(R⁴)R⁵, cyano, halo,nitro, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkynyl or substituted cycloalkynyl; Y is a straight or branchedchain C₁₋₆ alkylene group, cycloalkylene or substituted cycloalkylene;R¹ is hydrogen, alkyl, substituted alkyl, cycloalkyl or substitutedcycloalkyl; each R² independently is alkyl, substituted alkyl, alkoxy,substituted alkoxy, amino, substituted amino, aryl, substituted aryl,aryloxy, substituted aryloxy, cyano, cycloalkyl, substituted cycloalkyl,cycloalkoxy, substituted cycloalkoxy, heteroaryl, substitutedheteroaryl, heteroaryloxy, substituted heteroaryloxy, heterocyclyl,substituted heterocyclyl, heterocyclyloxy, substituted heterocyclyloxy,aminoacyl, aminoacyloxy, carboxyl, carboxyl ester, carbonate ester,—C(O)N(R⁴)R⁵, nitro or halo; each R³ independently is alkyl, substitutedalkyl, alkoxy, substituted alkoxy, amino, substituted amino, aryl,substituted aryl, aryloxy, substituted aryloxy, cyano, cycloalkyl,substituted cycloalkyl, cycloalkoxy, substituted cycloalkoxy,heteroaryl, substituted heteroaryl, heteroaryloxy, substitutedheteroaryloxy, heterocyclyl, substituted heterocyclyl, heterocyclyloxy,substituted heterocyclyloxy, aminoacyl, aminoacyloxy, carboxyl, carboxylester, carbonate ester, nitro or halo; each R⁴ independently ishydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl,heterocyclyl, substituted heterocyclyl, carboxyl, carboxyl ester, aryl,substituted aryl, heteroaryl, substituted heteroaryl, acyl or M⁺ whereinM⁺ is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄, wherein each R⁹ independently is hydrogenor alkyl, and the nitrogen immediately adjacent to R⁵ is anionic; or R⁴and R⁵ together with the intervening atom or atoms bound thereto, form aheterocyclyl or substituted heterocyclyl group; each R⁵ independently ishydrogen, alkyl, substituted alkyl, amino, cycloalkyl, substitutedcycloalkyl, heterocyclyl, substituted heterocyclyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, carboxyl, carboxyl ester oracyl; R⁷ is hydrogen, alkyl, substituted alkyl, cycloalkyl, substitutedcycloalkyl, heterocyclyl, substituted heterocyclyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl or acyl; R⁸ is hydrogen, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl or acyl; or R⁷ and R⁸ together, with nitrogen towhich they are bound, form a heterocyclyl or substituted heterocyclylgroup; R¹⁶ is a straight or branched chain C₁₋₆ alkylene group,cycloalkylene or substituted cycloalkylene; and each of R⁷ and R⁸optionally are M⁺ wherein M⁺ is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄, wherein each R⁹independently is hydrogen or alkyl, and the nitrogen bearing M⁺ isanionic.
 9. The compound of claim 8, wherein p is zero and each of R⁶and Y, independently, is a straight or branched chain C₁₋₆ alkylenegroup.
 10. The compound of claim 9, wherein each of R⁶ and Y,independently, is methylene or ethylene.
 11. The compound of claim 1,according to formula III:

wherein: p is 0, 1, 2, or 3; q is 0, 1, 2, or 3; X is alkyl, substitutedalkyl, hydroxy, alkoxy, substituted alkoxy, amino, substituted amino,carboxyl, carboxyl ester, —C(O)N(R⁴)R⁵, cyano, halo, nitro, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkynyl orsubstituted cycloalkynyl; each R² independently is alkyl, substitutedalkyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl,cycloalkoxy, substituted cycloalkoxy, heterocyclyl, substitutedheterocyclyl, heterocyclyloxy, substituted heterocyclyloxy, —C(O)N(R⁴)R⁵or halo; each R³ independently is alkyl, substituted alkyl, alkoxy,substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkoxy,substituted cycloalkoxy, heterocyclyl, substituted heterocyclyl,heterocyclyloxy, substituted heterocyclyloxy or halo; R⁶ is hydrogen,alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl,heterocyclyl, substituted heterocyclyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl or acyl; R⁷ is hydrogen, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl or acyl; R⁸ is hydrogen, alkyl, substitutedalkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, substituted heteroarylor acyl; or R⁷ and R⁸ together, with nitrogen to which they are bound,form a heterocyclyl or substituted heterocyclyl group; wherein at leastone of R⁷ and R⁸ is non-hydrogen; and each of R⁶, R⁷, and R⁸ optionallyare substituted with M⁺, wherein M⁺ is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄, whereineach R⁹ independently is hydrogen or alkyl, and the nitrogen bearing M⁺is anionic.
 12. The compound of claim 1, according to formula IV:

wherein: p is 0, 1, 2, or 3; q is 0, 1, 2, or 3; X is alkyl, substitutedalkyl, hydroxy, alkoxy, substituted alkoxy, amino, substituted amino,carboxyl, carboxyl ester, —C(O)N(R⁴)R⁵, cyano, halo, nitro, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkynyl orsubstituted cycloalkynyl; each R² independently is alkyl, substitutedalkyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl,cycloalkoxy, substituted cycloalkoxy, heterocyclyl, substitutedheterocyclyl, heterocyclyloxy, substituted heterocyclyloxy, —C(O)N(R⁴)R⁵or halo; each R³ independently is alkyl, substituted alkyl, alkoxy,substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkoxy,substituted cycloalkoxy, heterocyclyl, substituted heterocyclyl,heterocyclyloxy, substituted heterocyclyloxy or halo; R⁶ is hydrogen,alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl,heterocyclyl, substituted heterocyclyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl or acyl; R⁷ is hydrogen, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl or acyl; R⁸ is hydrogen, alkyl, substitutedalkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, substituted heteroarylor acyl; or R⁷ and R⁸ together, with nitrogen to which they are bound,form a heterocyclyl or substituted heterocyclyl group; wherein at leastone of R⁷ and R⁸ is non-hydrogen; and each of R⁶, R⁷, and R⁸ optionallyare substituted with M⁺, wherein M⁺ is K⁺, Na⁺, Li⁺ or ⁺N(R⁹)₄, whereineach R⁹ independently is hydrogen or alkyl, and the nitrogen bearing M⁺is anionic.
 13. A compound which is: I-1:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N,N-dimethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine;I-2:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N,N-dimethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine;I-3:N2-(3-Aminosulfonylphenyl)-N4-[4-[(N,N-dimethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine;I-4:N2-(4-Aminosulfonylphenyl)-N4-[4-[(N,N-dimethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine;I-5:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-ethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine;I-6:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-ethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine;I-7:N2-(3-Aminosulfonylphenyl)-N4-[4-[(N-ethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine;I-8:N2-(4-Aminosulfonylphenyl)-N4-[4-[(N-ethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine;I-9:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine;I-10:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine;I-11:N2-(3-Aminosulfonylphenyl)-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine;I-12:N2-(4-Aminosulfonylphenyl)-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine;I-13:N2-(3-Aminosulfonylphenyl)-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-5-fluoro-2,4-pyrimidinediamine;I-14:N2-(4-Aminosulfonylphenyl)-N4-[3-[(N-ethylsulfamoylamino)methyl]phenyl]-5-fluoro-2,4-pyrimidinediamine;I-15:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-ethylsulfamoylamino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine;I-16:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-ethylsulfamoylamino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine;I-17:N2-(4-Aminosulfonyl-3,5-dimethylphenyl)-5-chloro-N4-[4-[(N-ethylsulfamoylamino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine;I-18:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine;I-19:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]phenyl]-2,4-pyrimidinediamine;I-20:N2-(4-Aminosulfonyl-3,5-dimethylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoyl-amino)methyl]phenyl]-2,4-pyrimidinediamine;I-21:N2-(3-Aminosulfonylphenyl)-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine;I-22:N2-(4-Aminosulfonylphenyl)-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine;I-23:N2-(4-Aminosulfonyl-3,5-dimethylphenyl)-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]phenyl]-5-methyl-2,4-pyrimidinediamine;I-24:N2-(3-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine;I-25:N2-(4-Aminosulfonylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoylamino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine;I-26:N2-(4-Aminosulfonyl-3,5-dimethylphenyl)-5-chloro-N4-[4-[(N-cyclopropylsulfamoyl-amino)methyl]-2-methylphenyl]-2,4-pyrimidinediamine;I-27:N4-[3-(N-ethyl-N-propionyl)aminosulfonyl-(N-propionyl)aminomethyl]phenyl-5-methyl-N2-(3-propionylaminosulfonyl)phenyl-2,4-pyrimidinediamine;I-28:N4-[3-(N-ethyl-N-propionyl)aminosulfonyl-(N-propionyl)aminomethyl]phenyl-5-methyl-N2-(3-propionylaminosulfonyl)phenyl-2,4-pyrimidinediaminesodium salt; I-29: N2-(3-aminosulfonyl-4-methyl)phenyl-N4-(3-ethylaminosulfonylaminomethyl)phenyl-5-methyl-2,4-pyrimidinediamine;I-30:N2-(3-aminosulfonyl)phenyl-N4-(3-N,N-dimethylaminosulfonylaminomethyl)-5-methyl-2,4-pyrimidinediamine;I-31:N2-(4-aminosulfonyl)phenyl-N4-(3-N,N-dimethylaminosulfonylaminomethyl)-5-methyl-2,4-pyrimidinediamine;I-32:N2-(3-aminosulfonyl-4-methyl)phenyl-N4-(3-N,N-dimethylaminosulfonylamino-methyl)-5-methyl-2,4-pyrimidinediamine;II-1:N2-(3-aminosulfonyl-4-methyl)phenyl-N4-(2-N,N-dimethylaminosulfonyl-1,2,3,4-tetrahydroisoquinolin-7-yl)-5-fluoro-2,4-pyrimidinediamine;II-2:N2-(3-aminosulfonyl-4-methyl)phenyl-N4-(2-N,N-dimethylaminosulfonyl-2,3-dihydro-1H-isoindol-5-yl)-5-fluoro-2,4-pyrimidinediamine;II-3:N2-(4-aminosulfonyl)phenyl-N4-(2-N,N-dimethylaminosulfonyl-2,3-dihydro-1H-isoindol-5-yl)-5-methyl-2,4-pyrimidinediamine;II-4:N2-(3-aminosulfonyl)phenyl-N4-(2-N,N-dimethylaminosulfonyl-2,3-dihydro-1H-isoindol-5-yl)-5-methyl-2,4-pyrimidinediamine;or II-5:N2-(3-aminosulfonyl-4-methyl)phenyl-N4-(2-N,N-dimethylaminosulfonyl-2,3-dihydro-1H-isoindol-5-yl)-5-methyl-2,4-pyrimidinediamine.14. A pharmaceutical formulation comprising the compound according toclaim
 1. 15. A method of inhibiting an activity of a JAK kinase,comprising contacting the JAK kinase with an amount of the compound ofclaim 1 effective to inhibit an activity of the JAK kinase.
 16. A methodof inhibiting an activity of a JAK kinase, comprising contacting invitro a JAK3 kinase with an amount of the compound of claim 1 effectiveto inhibit an activity of the JAK3 kinase.
 17. A method of treating aT-cell mediated autoimmune disease, comprising administering to apatient suffering from such an autoimmune disease an amount of thecompound of claim 1 effective to treat the autoimmune disease.
 18. Themethod of claim 17, wherein the compound is administered in combinationwith, or adjunctively to, a compound or a pharmaceutical compositionthat inhibits Syk kinase with an IC₅₀ of less than 10 μM.
 19. A methodof treating or preventing allograft transplant rejection in a transplantrecipient, comprising administering to the transplant recipient anamount of the compound of claim 1 effective to treat or prevent therejection.
 20. The method of claim 19, wherein the compound isadministered to a tissue or an organ prior to transplanting the tissueor the organ in the transplant recipient.
 21. The method of claim 20,wherein the rejection is acute rejection.
 22. The method of claim 20,wherein the rejection is chronic rejection.
 23. The method of claim 20,wherein the rejection is mediated by HVGR or GVHR.
 24. The method ofclaim 20, wherein the allograft transplant is a kidney, a heart, a liveror a lung transplant.
 25. The method of claim 20, wherein the compoundis administered in combination with, or adjunctively to, anotherimmunosuppressant.
 26. The method of claim 25, wherein theimmunosuppressant is cyclosporine, tacrolimus, sirolimus, an inhibitorof IMPDH, mycophenolate, mycophanolate mofetil, an anti-T-Cell antibodyor OKT3.
 27. A method of treating or preventing a Type IVhypersensitivity reaction, comprising administering to a subject anamount of the compound of claim 1 effective to treat or prevent thehypersensitivity reaction.
 28. A method of treating a disease and/ordisorder of the eye, comprising administering to a subject an amount ofthe compound of claim 1 effective to treat the disease and/or disorderof the eye.
 29. The method of claim 28, wherein the disease and/ordisorder of the eye is dry eye syndrome, uveitis, allergicconjunctivitis, glaucoma or rosacea.